WO2010099607A1

WO2010099607A1 - Diagnostic test panel for the diagnostic of malaria and severe bacterial infections

Info

Publication number: WO2010099607A1
Application number: PCT/CA2010/000298
Authority: WO
Inventors: Susan Friedland; Kevin Kain
Original assignee: Fio Corporation
Priority date: 2009-03-02
Filing date: 2010-03-02
Publication date: 2010-09-10
Also published as: BRPI1009276A2; AP2011005906A0; AP3149A

Abstract

A test panel and kit, in package form, is for use in the diagnosis of malaria and severe bacterial infection in a test sample. They include monoclonal antibodies to histidine-rich protein H5 to P. falciparum aldolase and/or pan Pfalciparum lactate dehydrogenase, to Angiopoietin-1 and/or Angiopoietin-2, and to C-reactive protein and/or procalcitonin. A rapid diagnostic test device includes a portion to receive the panel, a chamber to receive the sample for contact with the monoclonal antibodies, and an element for detecting signals from the sample in contact with the monoclonal antibodies. Processors analyze the signals to differentiate between P.falciparum malaria likely or unlikely at risk for severe/cerebral malaria, non-P falciparum malaria, and infection which is likely or unlikely to be severe bacterial infection, in the sample. Also disclosed are an associated method and a computer readable medium which stores instructions for operating the test device.

Description

Diagnostic test panel for the diagnostic of malaria and severe bacterial infections

FIELD OF THE INVENTION

[0001] The present invention relates generally to the diagnosis of malaria and severe bacterial infection in test samples, and more particularly to a rapid diagnostic test panel, kit, device, method and computer readable medium for use in the diagnosis of malaria and severe bacterial infection in a test sample.

BACKGROUND OF THE INVENTION

[0002] Prior art malaria devices may typically have been useful only for performing single biomarker analyses. It may heretofore have been necessary to combine several results from several different diagnostic devices using various types of biological samples (e.g., whole blood, serum, plasma) to provide a doctor and/or biologist with the requisite information to perform a diagnosis and/or to make clinical management decisions.

[0003] Every year, malaria may be responsible for the deaths of about 800,000 children under five years of age in sub-Saharan Africa alone. Children under five, especially infants, may be particularly vulnerable to malaria due to a lack of immunity. Toddlers and older children may have partial immunity, which can complicate the identification of those who truly need treatment by attenuating clinical signs of infection. [0004] In sub-Saharan Africa, only a minority of children may have access to competent microscopy diagnosis. In endemic areas and/or in the absence of microscopy, the World Health Organization (WHO) case definition of malaria may be stated as the presence and/or a history of fever without obvious cause. (WHO, 1990). Yet there may be evidence that clinical diagnosis of malaria in resource-poor countries may be often extremely inaccurate. Attempts to come up with a predictive algorithm based on clinical signs and symptoms may not have been successful thus far. There may be little empirical evidence of the accuracy, predictability and/or reliability of using clinical signs and symptoms alone for diagnosing malaria within different endemic settings. Using physical exam and/or history alone to differentiate malaria from other causes of fever may heretofore have been notoriously difficult, because symptoms may be non-specific and/or may be shared by many other disease syndromes.

[0005] Microscopy may still be the "gold standard" laboratory diagnostic procedure for malaria in most locales. However, most children in sub-Saharan Africa may not have access to microscopy. Microscopy may involve skills that require extensive training and/or experience. What microscopy may be available may be of variable quality. Therefore many children may be admitted to hospital with a diagnosis of severe malaria when their symptoms may actually be due to other undiagnosed causes. [Reyburn H et al. (2004), "Overdiagnosis of malaria in patients with severe febrile illness in Tanzania: a prospective study". 5MJ 329(7476): 1212.] Antimalarial drugs may often be administered presumptively to febrile children. Lack of laboratory diagnosis with resultant reliance on clinical diagnosis, low-quality lab diagnosis, and/or widespread overtreatment may all contribute to a high pediatric mortality from malaria. Fear of rapid mortality from untreated malaria may be one of the drivers of presumptive treatment. There may, however, be increasing evidence that, if accurate diagnosis of malaria is available, febrile children (who have negative lab tests for malaria) may be safely treated without antimalarial drugs. [Hopkins H et al. "Effectiveness and safety of training in fever case management and malaria RDT use at health centers in Uganda". Abstract. 57^th ASTMH meeting, December 2008.] Extending the diagnostic capability to include rapid identification of probable severe bacterial illness may preferably, according to the present invention, increase the numbers of children who will appropriately receive antibiotics instead of antimalarial drugs.

[0006] Not only may inappropriate treatment have led to many deaths from non-malarial causes, but it may have also exacerbated the problem of antimalarial drug resistance. New drugs may be available (for example, artemisinin combined therapy or "ACT"), but they may be much more expensive. There may exist rapid diagnostic tests for malaria, but they may also be of variable quality, and they may not distinguish asymptomatic parasitemia from malaria illness. Because of the dramatically increased cost of treatment, accurate diagnosis may now be even more crucial for cost containment and/or for the health systems of developing countries (Rafael 2006).

Clinical Syndrome

[0007] Malaria may be generally thought to be caused by Plasmodium, a protozoan (single- celled) parasite. Four different Plasmodium species may be thought to cause human disease: i.e., P. vivax, P. malariae, P. ovale, and P. falciparum.

[0008] Malaria may have been characterized by periodic attacks (e.g., every about 2 to about 3 days, depending on species) of shaking chills, high fever, and/or sweating. [0009] Other indications may have included malaise, headache, arthralgias, hemoglobinuria, splenomegaly, leucopenia, and/or hemolytic anemia.

[0010] P. falciparum malarial infections may have caused severe complications: e.g., cerebral malaria with seizures, cerebral ischemia, coma, shock, renal/hepatic failure, metabolic acidosis, pulmonary edema, and DIC.

Pathophysiology

[0011] Malaria may have been generally thought to be transmitted from human to human by the female Anopheles mosquito. It may also have been transmitted by blood transfusion, and/or during pregnancy and/or birth.

[0012] In malarial infections, protozoa may have been generally thought to grow in the liver, then spreading to red blood cells where they reproduce. Red blood cells may have been generally thought to fill up with protozoa and burst at the same time (e.g., every about 48 hrs to about 72 hrs, depending on the species), releasing protozoa into the bloodstream and exposing them to immune system, resulting in about six hours of chills, fever, and/or sweats.

[0013] A severe complication of the disease may have been generally thought to result from sticky and/or burst red blood cells, which may block capillaries and/or venules, and/or obstruct blood delivery to organs and/or tissues. The process may have been previously categorized into chronic and acute versions. In case of falciparum, the process may result in rapid deterioration of health and/or in death. Differential diagnosis

[0014] The proper diagnosis of malaria may require various differential diagnoses, potentially including the following: influenza, bacteremia / sepsis, dengue fever, typhoid / paratyphoid, acute schistosomiasis, leptospirosis, yellow fever, East African trypanosomiasis (i.e., sleeping sickness), rickettsial fevers, and/or hepatitis.

Previous diagnostics

[0015] Clinical diagnosis may, by far, have been the most common method of diagnosis in resource-poor areas. People may have been presumed to have malaria on the basis of acute fever, +/- anemia and/or splenomegaly. Specificity of such diagnostics may be very low; especially insofar as the clinical presentation of malaria may be similar to that of other infections. Most cases of malaria in sub-Saharan Africa may have been previously diagnosed without the use of a laboratory.

[0016] Thick and thin blood smears of whole blood (which may have been stained and/or examined under a microscope for parasites) may heretofore have been the gold standard and mainstay of laboratory diagnosis. This type of test may still be the initial test for malaria in most of the world. It may have been used for initial diagnosis and/or for monitoring of the disease. Only a small percentage of malarial infections in resource-poor countries may have been confirmed by smear test, perhaps due in part to its low accessibility and/or availability in the prior art.

[0017] Smear tests may have been generally thought to afford a high sensitivity at relatively low cost per case, if performed correctly and with good equipment. Smear tests may heretofore have provided quantitative information. No sophisticated transport mechanisms may have been previously required for equipment related to the tests. Smear tests may have been found to be relatively inexpensive in high throughput situations.

[0018] The quality of smear tests may often have been poor - e.g., with sensitivities between about 50% and about 75%, and/or with specificities between about 59% and about 96%. Heretofore, smear tests may not have been widely available in rural areas. Smear tests may have previously required a trained technician, a microscope, electricity, and/or some consumables. Smear tests may become relatively expensive in low throughput situations.

[0019] Previously, lateral flow immuno-chromatographic antigen detection devices (i.e., a rapid diagnostic test or "RDT") may have been available, in the form of cassettes, dipsticks and/or cards. Such devices may have used blood from a finger prick. Fifteen minutes may have been required to provide a result using such prior art diagnostic tests. It may heretofore have been possible to detect one or more of the four following situations, via antigen capture: (a) detect only falciparum infection; (b) detect all species and distinguish falciparum from non- falciparum infection; (c) detect malaria infection without specifying the infecting species; and/or (d) detect only vivax infection.

[0020] Previously, there may also have existed antibody capture kits, reporting IgG and/or IgM to falciparum and/or vivax. Such kits may not have been generally used for case management.

[0021] Previous RDTs may have been somewhat more specific than, and/or about as sensitive as, good microscopy for falciparum. The sensitivity may have varied with the parasite density, the particular antibody used, and/or the shelf life. The cost may have been between about US $0.60 and about US $2.50. Previous RDTs useful for detecting falciparum, only, may have cost the least. Previous RDTs may have been relatively easy to learn to use, hardy, and/or cost-effective (as may be described in greater detail hereinbelow).

[0022] Previous Falciparum RDTs using HRP2 may have reported infection up to about 3 weeks after an actual infection may have been treated, potentially having led to false positives. Some RDTs may not have been adequately sensitive (or they may have been too sensitive in highly endemic areas, where many people may have parasitemia, but may not be ill). Previous RDTs may not have provided quantitative results. The cost of previous RDTs may have been too high for some settings. Some previous RDTs may have required cold chain transport, and/or some may have had only a relatively short shelf life. Cassette types of RDTs may have been are somewhat easier to use than strips, potentially therefore providing more reliable results, and/or being more cost-effective (even though they may have actually cost more).

[0023] Previous detection of antibodies may have been performed using serology - i.e., by indirect immuno-fluorescent analysis (IFA) and/or ELISA. Such detection systems, however, may not detect current infections, but instead only past infections. Similarly, serology may heretofore have required a lab setting.

[0024] Quantitative buffy-coat (QBC) centrifugal hematology may have been relatively easy to learn, and/or may have afforded a high sensitivity and/or specificity, while also being relatively quicker than smear tests, but it may also have been substantially equipment-dependent and, therefore, perhaps too expensive to be considered as an option for most settings.

[0025] PCR (polymerase chain reaction) may have afforded a very high sensitivity and/or specificity, but it may also have been substantially too equipment-dependent, complicated and/or expensive to be considered as an option for most settings in developing countries. PCT may have involved a time to result of > about 2 hrs. It may have been used, primarily in the developed world, to confirm the results of a blood smear test. Small PCR units which may require less lab infrastructure may be presently in development, but may not be widely available.

Prevention

[0026] Prior art means of malaria prevention may have involved one or more of the following: (i) prevention of mosquito bites with insecticide-treated nets (ITNs) over beds; (ii) prevention of mosquito bites with spraying of interior walls with DDT; (iii) prevention of mosquito bites with protective clothing; (iv) prophylactic anti-malarial medication.

[0027] Vaccines, genetically altered sterile mosquitoes and/or malaria-resistant mosquitoes may be presently in development.

Treatment

[0028] Perhaps the most common treatment for uncomplicated malaria of all species may be oral medication. Medications may prevent progression to severe forms of the disease, if effective and used properly. There may be a number of antimalarials presently available.

[0029] Effective treatment is, however, an exception rather than the rule, perhaps due in part to drug resistance, poor drug quality (i.e., there may presently exist a counterfeit malaria drug problem), poor compliance, and/or misdiagnosis.

[0030] Certain medications may be administered rectally and/or intramuscularly. In severe versions of the disease, intensive supporting treatment may be necessary. Treatment delays may be, and often are, fatal. Resistance may have developed to all classes of antimalarials, except perhaps to artemisinins.

[0031] To slow the development of resistance, artemisenin combined therapy ("ACT") may be used increasingly. Perhaps because the cost to provide treatment with ACT may be much higher than with older drugs, lab diagnosis may be cost effective.

Clinical course

[0032] The clinical course of the disease and/or its treatment may depend on many host variables (e.g., immune status, nutritional status, age, compliance, and/or genetic factors, such as, for example, sickle cell), treatment adequacy, species, patterns of drug resistance, and/or availability of hospitalization, if required. The course without treatment may be variable. Young children may be especially vulnerable to a severe course, perhaps due to a lack of immunity.

[0033] In very endemic areas, many adults may be infected, but they may not necessarily be ill, perhaps due to immunity. With prompt anti-malarial medication, prognosis of the disease may be generally good. In falciparum infections, when severe complications may develop, prognosis may be poor (between about 14% and about 17% mortality), perhaps even with treatment.

Malaria Epidemiology

[0034] The malaria life cycle may be complex, with the protozoa possibly taking one or more of several different forms in different environments. It may be generally believed that there may be three life cycles of the protozoa: one in the human liver; one in the human red blood cell; and one in the mosquito. Humans may have been considered to be intermediate hosts, and the female anopheles mosquito may have been considered to be the vector.

[0035] In terms of species distribution, P.vivax and P. falciparum may heretofore have been responsible for most infections in world. Falciparum may be the predominant species in Africa. Malariae may also be widely distributed, but may be somewhat less common. Vivax may be mostly uncommon in much of Africa; and in west Africa, it may be replaced mainly by ovale (which may be somewhat rare elsewhere). Vivax, although rarely fatal, may be more expensive to treat, because it may require treatment not only for the blood stage, but also for the liver stage.

[0036] Modes of transmission for malaria may be generally believed to include human to human transmission, and via a bite of the female anopheles mosquito. The incubation period for malaria may be generally believed to vary with the species of protozoan involved; and may usually be around two weeks (or between about 9 days and about 60 days).

[0037] The prevalence of malaria may be around one billion worldwide. There may be between about 300 and about 500 million cases per year worldwide, with more than about 1 million deaths per year (or between about 3000 and about 6000 deaths per day). There may be between about 200 and about 450 million cases of fever in children with parasites in Africa per year.

[0038] Malaria may be endemic in northern South America, in South and S.E. Asia, and/or in much of Africa, particularly, for example, in sub-Saharan Africa.

[0039] More than about 80% of malaria deaths may occur in sub-Saharan Africa, where about 66% of the population may be considered to be at risk. Most of the deaths may occur in African children under the age of 5 years, who may have no immunity and/or who may develop cerebral malaria. Less than about 15% of malaria deaths may occur in Asia and/or in Eastern Europe combined, despite about 40% of that population being considered to be at risk.

[0040] In the Americas, about 14% of the population may be considered to be at risk, but only a small fraction of the malaria deaths may actually occur there.

[0041] Recent malaria-related trends may have included one or more of the following: an increasing use of artemesinin combined therapy (ACT); an increasing drug and/or insecticide resistance, less so for ACT; an increased intensity in Southeast Asia; a re-emergence in Central Asia and/or in Trans-Caucasian countries; an improved coordination and scaling up of programs worldwide since 2000; an increasing risk in the developed world, perhaps due to travel, global climate changes and/or antimalarial resistance; malaria may be the most common cause of febrile illness without localizing signs in the returning traveler, although dengue may surpass it in travelers returning from Asia and/or the Caribbean; recent significant progress in the development of malaria vaccines, and/or in the development of sterile Anopheles mosquitoes; and/or significant progress in control by using insecticide treated nets (ITNs) on beds.

[0042] Recent constraints on malaria control may have included one or more of the following: lack of access to proper diagnosis and/or treatment; lack of health infrastructures; lack of effective malaria control policies; drug resistance, which may have caused between about a 4-fold and about an 8-fold estimated increase in mortality; insecticide resistance; and/or lack of effective vaccines. Socioeconomic burden

[0043] Malaria itself may have been thought to have caused great economic disability and/or instability. It may have been generally believed to have incapacitated (and/or to have caused incapacitation of) the work force, to have lowered educational achievement, and/or to have discouraged tourism and/or business investment. It may have prevented some countries from mounting effective malaria control policies, potentially having thereby set up a vicious cycle. In Africa, malaria may be believed to account for as much as about 40% of public health expenditures, between about 30% and about 50% of hospital admissions, and/or about 50% of outpatient visits. In some African countries, it may be generally thought that reductions in the GDP by as much as about 30% may have been attributable to malaria. Malaria may have been the cause of immense opportunity loss, which may have been measured at about 45 million DALYs (perhaps about 13% of the total infectious disease contribution).

Potential Clinical Impact

[0044] It may be generally believed, though it is not essential to the present invention, that an improved malaria diagnostic method and/or system may have a large favourable impact on the otherwise high paediatric mortality from malaria. A recent model may have been constructed to demonstrate the attributable benefits of a new diagnostic system and/or method that may replace clinical judgement. Though not essential to the working of the present invention, such model may have indicated that diagnostic tests requiring minimal, or no, infrastructure may produce the largest gains in < 5 year old adjusted lives saved (i.e., as may be adjusted for the harm associated with treatment, plus the lives which may be saved through reduction of over-treatment), in malaria-related deaths averted, and/or in unnecessary treatments averted. Again, though not essential to the working of the present invention, the group responsible for having constructed this model may also have recommended the adoption and/or development of a new, undisclosed and/or hypothetical malaria test of a type which might have: (a) minimal infrastructure requirements, > about 95% sensitivity, and from about 90 to about 95% specificity, so as to potentially save an estimated 1,798,532 adjusted lives in one year; or (b) no infrastructure requirements, about 90% sensitivity, and about 90% specificity, so as to potentially save an estimated 2,192,062 adjusted lives in one year.

[0045] Though not essential to the working of the present invention, a report (e.g., Rafael, 2006) on this model may have assumed that adequate medication is available.

Further Background

[0046] Though not essential to the working of the present invention, it may be believed [see, for example, Rafael 2006; and Reyburn, H. et al. (2004), "Overdiagnosis of malaria in patients with severe febrile illness in Tanzania: a prospective study". BMJ 329(7476): 1212] that there is a large amount of misdiagnosis in malaria. This misdiagnosis may be believed to be due to difficulty in differentiating malaria from other causes of non-localizing fever, widespread lack of access to lab diagnosis, low quality microscopy, and/or ineffective clinical diagnosis algorithms. Accordingly, persons having ordinary skill in the art may appreciate a significant need (to exist) for a new malaria panel diagnostic device, system and method, particularly (for example) in malaria-endemic areas.

[0047] Although CRP may be a good indicator of infection and/or inflammation [perhaps even significantly better than the white blood cell (WBC) count which may heretofore have been typically used], it may not accurately discriminate bacterial infections from viral ones. While CRP levels for bacterial infections may tend to be > about 40 mg/L, and for viral infection it may tend to be lower, there may be significant overlap. High CRP levels may be seen in association with adenovirus, CMV, influenza, measles, mumps and/or other viral infections. [See, for example, Hansson, L.O. et al (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25:625-630; and Clarke, D. et al (1983), "Use of serum C- reactive protein in differentiating septic from aseptic meningitis in children". J Pediatr 1983, 102:718-720.] CRP may also afford less than ideal discrimination between malarial infections and bacterial ones [see, for example, Ericksson, B. (1989), "Changes in erythrocyte sedimentation rate, CRP and hematological parameters in patients with acute malaria". Scand J Infect Dis 21(4): 4334-41]. Accordingly, persons having ordinary skill in the art may appreciate a disadvantage in establishing an inflexible cut-off value for diagnoses based on CRP levels. According to one aspect of the present invention, CRP results may be reported quantitatively and/or semi-quantitatively, so as to provide a user with additional data concerning CRP levels.

[0048] Though not essential to the working of the present invention, CRP levels may be generally believed to vary widely between individuals, by age, by pathogen, by locale and/or according to other host factors. Receiver operating curve (ROC) analyses may heretofore have been generally used to determine cut-off points between normal and abnormal CRP levels. In the prior art, cut-off points may have been somewhat variable, perhaps due in part to differences in study populations. For example, one prior art study may have indicated community paediatric norms in a Tanzanian locale with high malaria endemicity to be about twice that of a European norm. Accordingly, persons having ordinary skill in the art may appreciate a disadvantage in establishing an inflexible cut-off values. According to one aspect of the present invention, local normative data may be gathered prior to establishing cut-off values. [0049] With regard to the use of CRP in work-ups of undifferentiated paediatric fevers in North America, CRP may not heretofore have been typically used in the work-ups of non- localizing fevers in young children, even though WBC count (which may still be widely used) may be less accurate. CRP may heretofore have been the subject of some controversy in American paediatric literature, which may have presented a range of opinions. In one camp are those who may consider CRP to be a useful tool for investigating the cause of a fever [e.g., Pulliam, P. et al (2001), "CRP in febrile children 1 to 36 months of age with clinically undetectable serious bacterial infection". Pediatrics 108: 1275-1279], especially when the duration of the fever may be longer than about 12 hrs [e.g., Pratt, A. et al. (2007), "Duration of fever and markers of serious bacterial infection in young febrile children". Peds Ml 49: 31-35]. Those in this first camp may not, however, consider CRP to be appropriate for use as a single test. In another camp are those who may have concluded that, even though CRP may perform better than ANC and/or WBC as a screen for occult bacterial infection, it may offer little help to a clinician evaluating a febrile child. [See, for example, Isaacman (2002), "Utility of the Serum CRP for Detection of Occult Bacterial Infection in Children". Arch Pediatr Adolesc Med 156: 905-90; Carrol, E. D. et al. (2002), "Procalcitonin as a diagnostic marker of meningococcal disease in children presenting with fever and a rash". Arch Dis Child 85: 282-285; and Kupperman, N. (2002), "The Evaluation of Young Febrile Children for Occult Bacteremia". Arch Pediatr Adolesc Med 156:855-857.] In North American paediatrics, CRP may heretofore have been more commonly used to monitor patient response to treatment for chronic infections and/or inflammatory conditions - such as, for example, osteomyelitis, juvenile rheumatoid arthritis (JRA) and Crohn's Disease (Jaye 1997). One exemplary situation involving acute infection in which CRP may heretofore have been used may have been in neonatal sepsis workups. Though not essential to the working of the present invention, serial CRP tests on Day 1 and 2 may have been generally thought to afford a very high negative predictive value [Benitz et al. (1998), "Serial C-Reactive Protein Levels in the Diagnosis of Neonatal Infection". Pediatrics 102, e41, 1998]. Though not essential to the working of the present invention, persons having ordinary skill in the art may appreciate CRP to have been previously used in North American centers for the aforesaid purpose [Polen, R. (2005), "Neonatal Sepsis". NICUniversity On-line CME Course]. There may exist a gap between persons having ordinary skill in actual clinical practice and in the existing knowledge base, especially in North America. Though not essential to the working of the present invention, there may exist a need to educate general paediatricians concerning the potential utilities of rapid point-of-care CRP tests. Neonatology and rheumatology specialists having ordinary skill in the art, however, may appreciate (there to exist) a need for rapid point-of-care CRP tests.

[0050] Most of the prior art studies on the use of CRP in the work-up of paediatric undifferentiated fevers may originate from Europe, where the use of CRP may be viewed somewhat more favourably. Though not essential to the working of the present invention, these studies may have been generally believed to indicate that CRP may have performed somewhat better than WBC and/or ANC as a predictor of severe bacterial illness, and/or that CRP may be reasonably included as part of an associated work-up [see, for example, Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric InfDis J 18: 875-881; Galetto-Lacour 2000; Hsiao, A. et al. (2005), "Fever in the new millennium: a review of recent studies of markers of serious bacterial infection in febrile children". Curr Opin Peds 17: 56-61; and Andreola, B. (2007), "Procalcitonin and CRP as Diagnostic Markers of Severe Bacterial Infections in Febrile Infants and Children in the Emergency Department". Pediatr Infect DIs J 26(8): 672-677]. In Europe, there may be somewhat more clinical use of CRP in such work-ups. It may be somewhat difficult to generalize the data from prior art studies, perhaps due in part to differences in case definitions and/or in cut-off values. As well, there may be wide variability in sensitivities and/or specificities - e.g., from about the mid 60s to about the low 90s. Persons having ordinary skill in the art may appreciate there to exist a need in Europe for rapid point-of-care CRP and/or PCT tests, which may heretofore have been left unsatisfied by prior art products.

[0051] In the prior art pertaining to adults, CRP may heretofore have found somewhat more widespread application for monitoring post-op infections, for identifying sepsis in the ICU, for severe burns, and/or for auto-immune diseases. Burn unit and ICU specialists having ordinary skill in the art may appreciate there to exist a need for rapid point-of-care CRP tests.

[0052] Prior art data on the existence and extent of a potential relationship between CRP and malaria may be scant, somewhat contradictory, and/or not reflective of current conditions. Some prior art data may indicate a relationship between elevated CRP and parasite density, which may be attenuated in semi-immune individuals. Other prior art studies - e.g., a study conducted in an endemic area - may indicate CRP levels to substantially correlate with acute malaria episodes only in babies under 12 months. It may have been generally thought (though it is not essential to the working of the present invention) that, in babies under 12 months, high CRP levels may be caused by malaria about 80% of the time. It may also be generally believed (though, again, it is not essential to the working of the present invention) that, by about 3 to 5 years of age, high CRP levels may be attributable to malaria only about 20% of the time, and/or that any earlier relationship between parasite density and CRP may be lost. It may have been believed that this change may have been due, perhaps in part, to development of a semi-immunity during the second and third years of life. Some of the prior art may have suggested that, in endemic areas, a high CRP level may be likely to indicate acute malaria, whereas in older infants and toddlers a high CRP may be thought more likely to indicate a non-malaria acute infection [Hurt, N. (1994), "Do High Levels of CRP in Tanzanian Children Indicate Malaria Morbidity?" CHn Diag Lab Immunol, July 1994: 437-444]. According to one aspect of the present invention, the CRP result may be most helpful in the case of a negative malaria result, and/or in the case of a positive malaria result, the specificity of a positive CRP result may be reduced.

[0053] Procalcitonin (PCT) may be a host biomarker. It may heretofore have been generally believed, though it is not essential to the working of the present invention, that PCT may have a greater accuracy than CRP as a diagnostic marker of severe bacterial infection, and/or that it may be a better discriminator between bacterial infections and viral ones in children, in adults, or in both children and adults. A prior art meta-analysis of fifteen studies may have indicated PCT to be superior, over CRP, in diagnosing sepsis in critically ill adults [Uzzan, B. et al. (2006), "Serum PCT in uncomplicated falciparum malaria. A preliminary study". Trav Med & lnf Dis March 2006, pp 77-80]. It may be generally believed, though it is not essential to the working of the present invention, that such a result is also consistent with the results of most prior art paediatric studies - see, for example, Dubos F (2006), "Serum PCT and other biologic markers to distinguish between bacterial and aseptic meningitis". J Pediatr 149 (72-6); Hansson, L. O. et al. (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25: 625-630; Hsiao, A. et al. (2005), "Fever in the new millennium: a review of recent studies of markers of serious bacterial infection in febrile children". Curr Opin Peds 17: 56-61; Galetto-Lacour, A. (2001), "Precalcitonin, IL_6, IL-8, IL-I receptor antagonist and CRP as identificators of serious bacterial infections in children with fever without localising signs". Eur J Pediatr 160: 95-100; and Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric Inf Dis J 18: 875-881.

[0054] It may be generally believed (though it is not essential to the working of the present invention) that PCT levels may rise in the setting of P. falciparum, but perhaps that they may not rise universally. Prior art studies may have indicated that PCT levels may not rise in the setting of P.vivax [e.g., Manegold et al. (2003), "PCT levels in tertian malaria". Malar J 16, 2:34. Epub 2003 Oct 16]. In the prior art, high PCT concentrations may heretofore have been highly correlated with a lack of semi-immunity (e.g., Chiwakata 2000, and Uzzan 2005).

[0055] In the prior art, a few researchers may have suggested combining the results of CRP and PCT tests to increase sensitivity and/or specificity [Ip, M. et al. (2007), "Value of serum procalcitonin, neopterin, and CRP in differentiating bacterial from viral etiologies in patients presenting with lowere respiratory tract infections". Diagn Microbiol & Inf Dis 59: 131-136]. In one exemplary prior art study, a high PCT level coexisting with a high CRP level may have been generally believed to indicate a systemic infection, while a low PCT level coexisting with a high CRP level may have been generally believed to indicate an acute phase response or local infection, but no systemic infection [Rothenburger M et al. (1999), "Detection of Acute Phase Response and Infection. The Role of Procalcitonin and CRP". Clin Chem and Lab Med 37(3): 275-279]. According to one aspect of the present invention, such indications may be further investigated, confirmed and/or relied upon in operation of the device, system and/or method - particularly in situations where according to one aspect of the present invention, the device, system and/or method may report the PCT and CRP results dichotomously. [0056] In Europe, the prior art may include point-of-care (POC) rapid diagnostic tests for CRP and/or PCT levels.

[0057] CRP tests may not be currently used, on any widespread basis, in the assessment of non-localizing severe febrile illness in North America. Neither CRP nor procalcitonin (PCT) may be currently used in paediatric septic work-ups apart, potentially, from some limited use in neonatal work-ups. CRP tests may have been previously used in paediatrics, but their principal use(s) may have been in following subacute and/or chronic conditions such as, for example, osteomyelitis and/or inflammatory bowel disease (and then mainly as a substitute for sedimentation rate tests). Procalcitonin may have been hardly ever used in North America apart, potentially, from some very limited use in neonatology.

[0058] While the potential benefits may be staggering in their favourable applications, there may be little perceived need among paediatricians for rapid diagnostic malaria tests, CRP tests, and/or PCT tests. Though not essential to the working of the present invention, it may be appreciated by persons having ordinary skill in the art that neonatologists, clinicians in malaria endemic areas, and/or European clinicians may perceive a substantial need for one or more such tests and/or for improved versions of same.

Prior Art Difficulties in Differentiating Malaria from Other Causes of Fever

[0059] The differentiation of malaria from other causes of fever in the absence of microscopy may heretofore have been perceived to be difficult.

[0060] In one exemplary prior art sample of Philippine children under age 9 which may have been conducted in an endemic area, only about 52% of fever cases may have had parasitemia, potentially indicating that the WHO's recommended case definition of malaria may not be adequately specific [Gomes, M. et al. (1994), "Symptomatic identification of malaria in the home and in the primary health care clinic". Bull World Health Otgan 72(3): 383-90]. In a community-based cohort of about 561 non-HIV+ children in and around Kampala, aged 1 to 1 1 years, which may have been followed for about 1 1 months, about 33% of the febrile episodes may have been accounted for by malaria [Kamya, M. et al. (2007), "Effects of TMP-SMZ and insecticide-treated bednets on malaria among HIV-infected Ugandan children". AIDS, 21(15): 2059-66].

[0061] There may have been several attempts to develop a better clinical algorithm. One example may be a study of about 1527 Thai children, aged 2 to 15 years, in which clinical predictors of malaria (e.g., confirmed fever, headache, joint pain, absence of diarrhea, splenomegaly, hepatomegaly, clinical anemia, and/or absence of cough) may have been combined in numerous ways. Even when combined into the best diagnostic algorithm, such an algorithm may have resulted in the erroneous treatment of malaria in about 29% of non-malaria episodes, and may have left untreated about 51% of the true malaria cases [Luxemburger, C. et al. (1998), "Clinical features cannot predict a diagnosis of malaria or differentiate the infecting species in children living in an area of low transmission". Trans R Soc Trop Med Hy g. 92(1): 45- 9]. Also, in the prior art, manualized diagnostic protocols using clinical signs often may have been not followed [Ray, S. (1995), "Clinical audit of malaria diagnosis in urban primary curative care clinics, Zimbabwe". Cent Afr J Med 4\(\2): 385-91].

[0062] In the prior art, clinical diagnosis of malaria may have resulted in over-diagnosis in certain locales. In about 688 Tanzanian children, at a rural government clinic in Tanzania (in an area of low endemicity), presumptive treatment based on clinical diagnosis may have resulted in a substantial proportion of the children being inappropriately managed - i.e., from about 39% to about 88%, depending on the method of malaria case definition [Massaga J (1999), "Malaria presumptive treatment in Tanzania: is it rational approach for malaria management in rural health units?" Afr J Health ScL 6(1): 22-6]. At a Togo clinic, most patients later diagnosed with relapsing fever may have been treated for malaria, regardless of whether plasmodia were observed - possibly having resulted in ineffective treatment for RF [Nordstrand, A. et al. (2007), "Tickborne relasing fever diagnosis obscured by malaria, Togo". Emerg Infect Dis 13(1): 1 17- 23]. Invasive salmonellosis may be common in tropical Africa, and/or may typically present itself as a non-specific febrile illness that may be difficult to distinguish clinically from malaria. At a mission hospital in rural Zaire, about 62% of children aged 1 to 15 years with prolonged fever, with no focus on PE, and with negative or scanty thick film for malaria may have had culture and/or serology-proven salmonella [Cheesbrough J et al. (1997), "Clinical definition for invasive Salmonella infection in African children". Pediatr Infect Dis J. 16(3): 227-83].

[0063] Malaria may often be missed as well. In a teaching hospital in Ibadan, in a series of about 16 Nigerian adolescents which may have been presenting with fever and/or altered consciousness, the diagnosis of cerebral malaria may have been initially considered for only one adolescent. None of these adolescents may have received microscopy for malaria. The most common erroneous diagnoses may have been typhoid (7), meningitis (6) and/or encephalitis (2). [Sowunmi 1994.]

[0064] Malaria may also have been often missed in the U.S. emergency department settings. Imported falciparum malaria presenting to emergency departments in the United States may have been frequently misdiagnosed. Of twenty cases of P .falciparum identified at a Los Angeles medical center, malaria may have been considered in the ED in only about 60% of the cases, and P. falciparum may have been considered in only about 2 cases (10%). [Kyriacou, D. (1996), "Emergency Department Presentation and Misdiagnosis of Imported Falciparum Malaria". Annals Emerg Med, 27(6): 696-699.] At an Atlanta teaching hospital, the diagnosis may have been missed in about 71% of about 126 cases [Vicas, A. (2005), "Imported malaria at an inner- city hospital in the United States". Am L Med Sci 329(1): 6-12]. With only about 1528 malaria cases in 2005 [Thwing, J. et al. (2007), "Malaria surveillance - United States, 2005". MMWR Surveill Summ 56(6): 23-40], for example, there may not have been not much reinforcement in the U.S. for including malaria in the differential diagnosis, although this may change with an increased incidence of malaria.

C-Reactive Protein in the Prior Art

[0065] C-reactive protein (CRP) may be a plasma protein which may be produced by the liver. CRP may provide a simple non-specific measure of inflammation, disease severity and/or progression, efficacy of therapy, and/or severity of complications. It may be generally believed, though it is not essential to the working of the present invention, that CRP may not be used as a single diagnostic tool. CRP may, however, be diagnostically helpful in combination with other labs and/or clinical symptoms and/or signs, and it may be used in infectious disease, cardiology, rheumatology and/or transplantation medicine.

[0066] CRP may be generally believed to be a member of the class of acute phase reactants, which may rise dramatically during inflammatory processes in the body. The liver may release CRP in response to elevated levels of the cytokines, particularly IL-6. CRP may bind to phosphoryl choline on microbes. It may be believed, though it is not essential to the working of the present invention, that CRP may assist in binding complement to foreign and/or damaged cells, and/or that CRP may enhance phagocytosis by macrophages, which express a receptor for CRP.

[0067] CRP may be produced within about 4 hours to about 6 hours after onset of tissue injury and/or inflammation. Its serum level may double every about 8 hours, before peaking at about 36 hours to about 50 hours. With a very intense stimulus, its concentration may increase about 1000 fold, to levels above about 500 mg/L. After removal of the stimulus, CRP may fall rapidly, as it may have a half-life of about 19 hrs, and/or elevated CRP levels may return to about baseline within about a week [Koj, A. (1985). "Catabolism and turnover of acute phase proteins", p 145-160 in AH Gordon and A Koj (ed), The acute phase response to injury and infection. Elsevier, Amsterdam]. However, CRP may remain elevated if the underlying cause of the elevation persists. One single determinant of CRP level may be its rate of synthesis, which may in turn depend on the inflammatory insult intensity. With the potential exception of severe hepatic failure, CRP may rise whenever an inflammatory process may be present.

[0068] CRP elevations may be seen with most invasive infections. Gram positive and/or negative bacterial infections and/or fungal infections may cause marked CRP rises - potentially even in immunodeficient patients. By contrast, CRP increases may tend to be more modest in viral infections. This rule, however, may not be absolute. Uncomplicated infections with adenovirus, measles, mumps, and/or influenza may sometimes be associated with high CRP levels. Systemic viral infections with CMV and/or herpes simplex may also induce marked increases, as may parasitic diseases such as malaria, Pneumocystis and/or toxoplasmosis. TB and/or leprosy may cause only modest elevations. Non-infectious conditions that may also cause substantial changes may include trauma, surgery, auto-immune disease, and/or other inflammatory conditions, such as, for example, graft versus host disease. [Povoa, P. (2002), "CRP: a valuable marker of sepsis". Int Care Med DOI 10.1007/sOO 134-002- 1209-6 (online journal).]

[0069] The average concentration of CRP in healthy U.S. adult populations may be under about 1 mg/L, with a median of about 0.8 mg/L, and may be below about 10 mg/L in about 99% of normal samples. Levels between about 1 mg/L and about 10 mg/L may be correlated with an increasing risk of cardiovascular disease in adults, with levels of > about 3 mg/L potentially indicating a high risk. Levels higher than about 10 mg/L may indicate active acute and/or chronic inflammation and/or infection. US reference ranges from the National Health and Nutrition Examination Survey, 1999-2000 for children may be as follows: ages 3-9 yo, 1.4 mg/L; ages 10-15 yo, 1.6 mg/L; ages 16-19 yo, 1.8 mg/L. The same statistics, excluding children with CRP > 10 mg/L, may be as follows: ages 3-9 yo, 0.8 mg/L; ages 10-15, 0.9 mg/L; ages 16-19 yo, 1.4 mg/L. [From Ford, E. et άl. (2003), "C-Reactive Protein: US reference ranges from the National health and Nutrition Examination Survey, 1999-2000" Clin Chem, 49 (1353- 1357).] Laboratories may differ in the clinical cut-offs used. The typical clinical cut-off may have been 10 mg/L, although some labs may have used 5 mg/L as the clinical cut-off. The acute range may be between about 20 mg/L and about 500 mg/L. The appropriate assay range may be about 5 mg/L to the upper range of the assay.

[0070] Prior art analytical methods which have been available for assessing CRP levels may have included ELISA, immunoturbidimetry, rapid immunodiffusion and/or visual agglutination. There may be several rapid diagnostic tests (< about 5 min) for CRP, potentially including: (i) a three-minute turbidimetric quantitative test, e.g., the QuickRead CRP whole blood/serum/plasma assay (such as may have been offered by Orion Diagnostica, of Finland) which may have used a cut-off of about 8 mg/L, and (ii) a two-minute reflectometric quantitative test, e.g., the Nycocard CRP Whole Blood Test (such as may have been offered by Axis-Shield, of Norway). There may have been numerous prior art validation studies which may have been conducted with variable results. The QuickRead test may have been shown, in a purportedly independent study, to give the same quantitative results as those obtained using a reference laboratory method [Esposito, S. (2005), "Evaluation of a rapid bedside test for the quantitative determination of CRP". Clin Chem and Lab Med 53(4): 438-440]. Similarly, the NycoCard test's correlation coefficient - when compared to a reference method - may have been about 0.94 [Dahler-Ericksen, B. et al. (1997), "Evaluation of a near-patient test of CRP used in daily routine in primary healthcare by use of different plots". CUn Chem 43: 2064-2075]. The minimum detection limits, however, may be high - perhaps about 10 mg/L for the NycoCard test, and perhaps about 8 mg/L for the QuikRead test - and/or the measurement ranges may cover less than two orders of magnitude.

Prior Art uses of CRP in Malaria Endemic Areas

[0071] CRP levels may rise in many types of infections, including malaria. In the absence of parasitemia, a very high CRP level may suggest bacterial and/or viral infection, but it may not rule out other causes of inflammation. In the presence of parasitemia, a high CRP level may suggest a clinical malaria episode, but it may not rule out a concurrent non-malarial infection. CRP levels which may have been previously seen in bacterial infections, viral infections and/or malaria episodes may overlap in magnitude.

[0072] CRP levels may vary widely by age, by pathogen, by locale and by other host factors. Normal levels of CRP may not be not well-defined in developing countries. Increases in CRP levels, which may be associated with any given trigger, may vary widely among individuals. CRP results may be most meaningful when one may be able to provide local normative data stratified by age. According to one aspect of the present invention, local normative data stratified by age may be gathered over time.

[0073] Very limited data in the prior art may suggest that median CRP levels in African community samples may be elevated above European norms. In one prior art study, a median CRP level among Tanzanian children may have been about 6 mg/L, compared to about 3 mg/L in a European community sample. This CRP may have been measured against a background of increased inflammation from numerous causes. [Hurt, N. (1994), "Do High Levels of CRP in Tanzanian Children Indicate Malaria Morbidity?" Clin Diag Lab Immunol, July 1994: 437-444; and Naik, P. et al. (1984), "Serum CRP levels and falciparum malaria". Trans R Soc Trop Med //yg 78: 812-813.]

[0074] In endemic areas, the point prevalence of parasitemia among children may be very high - in one prior art study, about 87% of children under 6 yo. In the same study, the point- prevalence of CRP > about 6 mg/L may have been about 52%. Of those children with CRP > about 6 mg/L, about 27% of the elevated CRP levels may have been attributable to malaria. About 14% of all children may have had a CRP > about 6 mg/L attributable to malaria. The point prevalence of malaria-attributable fever may have varied between about 4% in infants to about 2% in young children. [Hurt, N. (1994), "Do High Levels of CRP in Tanzanian Children Indicate Malaria Morbidity?" Clin Diag Lab Immunol, July 1994: 437-444.]

[0075] In the prior art, it may have been thought that the proportion of high CRP levels attributable to malaria in children may have decreased quickly with age. During the first year of life, very high CRP levels may have been attributed to malaria about 82% of the time, but by about age 3 to about age 5, high CRP levels may have been attributable to malaria only about 20% of the time. The prior art may have presumed that other causes may have accounted for the other about 80% of high CRP levels. Regardless of how ill the child may have appeared, high malaria density may have been highly correlated to elevated CRP in the first year of life, but the relationship may have disappeared quickly over the first five years, as children may have started to develop immunity. By the time children were about 2 yo, there may have been no relationship between parasite density and CRP. Also, babies with a malaria episode may most commonly have had CRP levels > about 40 mg/L, but older children with a malaria episode may most often have had CRP levels between about 6 mg/L and about 10 mg/L. [Hurt, N. (1994), "Do High Levels of CRP in Tanzanian Children Indicate Malaria Morbidity?" CHn Diag Lab Immunol, July 1994: 437-444.] The tables shown in Figures 4 and 5 may have been reproduced, in whole or in part, from the same study.

[0076] A prior art study of asymptomatic children in Papua New Guinea with parasitemia may have helped to corroborate a thesis that children may modulate inflammation associated with parasitemia. The prevalence of CRP levels which may be consistent with an acute phase response (> about 10 mg/L) may have been relatively low (< about 10%). [Imrie, H. (2007), "Low Prevalence of an Acute Phase Response in Asymptomatic Children from a Malaria- Endemic Area of Papua New Guinea". AMJ Trop Med Hyg 76(2): 280-284.]

[0077] By adulthood, a CRP response to malaria may also be somewhat modulated. In a study [e.g., Graninger, W. et al. (1992), "Serum protein concentration in P .falciparum malaria". Act Trop 52: 121-128] of CRP levels in about 37 adult Brazilian patients, which may have had uncomplicated P. falciparum malaria, modest increases in CRP may have been found, as shown in the table reproduced in Figure 6 (grouped by history into non-immune and semi-immune groups). Prior Art uses for Malaria Diagnosis in Returning Travelers

[0078] High CRP levels (with a mean of about 49 mg/L), which may have been measured in adult returning travelers with acute uncomplicated malaria, may have been previously shown to return to about baseline within about one week of treatment. (Gillespie 1991.)

[0079] In the prior art, CRP levels may not have discriminated between malaria and non- malarial causes of fever among returning travelers; an average CRP may have been about the same in both groups [potentially including, for example, Ericksson, B. (1989), "Changes in erythrocyte sedimentation rate, CRP and hematological parameters in patients with acute malaria". Scand J Infect Dis 21(4): 4334-41].

Use to Predict Severe Bacterial Infection in Children

[0080] Since about the time of the introduction of the Hib vaccine and/or the pneumococcal vaccine in North America, the incidence of occult bacteremia among highly febrile infants and/or young children may have dropped dramatically. There may have been an estimated about 89% to about 94% drop in pneumococcal disease. [Black, S. et al. (2001), "Postlicensure evaluation of the effectivenesss of seven valent pneumococcal conjugate vaccine". Pediatr Infect Dis J 2001, 20: 1105-1 107.] In the prior art, there may have been some debate as to whether the routine practice of obtaining blood cultures and/or CBCs in previously healthy well-appearing highly febrile children from between about 3 mos and about 36 mos may still be indicated. [Kupperman, N. (2002), "The Evaluation of Young Febrile Children for Occult Bacteremia". Arch Pediatr Adolesc Med 156: 855-857.] Although blood culture may still have been the gold standard for diagnosing occult bacteremia in children, results may have been delayed, necessitating the expense and/or risk of hospitalization. Recently, the standard of care may have shifted toward more frequent assumptions that fever may have viral causes, and/or toward higher thresholds for full septic workups and/or hospital admissions for children over 2 months with non-localizing fever. For children under about two months, the threshold for septic work-up and/or admission may remain low. [Ishimine P (2006), "Fever without source in children 0 to 36 months of age". Ped Clin NA 53(2): 167-194.]

[0081] In the prior art, two biomarkers may have been most commonly studied as markers of severe bacterial infection (SBI) in children: CRP and procalcitonin. CRP may have been thought to be a better predictor of SBI in children than WBC and/or ANC, both of which may have been widely used, especially in infants, and/or especially when fever duration may have been longer than about 12 hrs. The American prior art may have mixed this use of CRP in combination with other lab, history and/or findings on PE.

[0082] WBC may have been previously judged to be the least sensitive and/or specific test in children, as compared to PCT, CRP and/or interleukin-6. PCT and/or CRP may have been considered two useful tools for investigating cause of fever in infants and/or children, but may not have been considered appropriate for use as a single test. [Hsiao, A. et al. (2005), "Fever in the new millennium: a review of recent studies of markers of serious bacterial infection in febrile children". Curr Opin Peds 17: 56-61.]

[0083] Although CRP may have been thought to be a good indicator of infection and/or inflammation, it may not have been regarded as a good discriminator of viral infections in comparison to bacteria ones. While CRP levels in bacterial infections may have tended to be > about 40 mg/L, and < about 40 mg/L in viral infections, this distinction may not have been regarded as absolute, and high CRPs may have been seen in adenovirus, CMV, influenza, measles, mumps and/or other viruses. [See, for example, Hanssen, L. O. et al. (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25: 625-630; and Clark, D. et al. (1983), "Use of serum C-reactive protein in differentiating septic from aseptic meningitis in children". J Pediatr 1983, 102: 718-720.] CRP may also have been regarded as non-discriminatory between malaria and bacterial infections [Ericksson, B. (1989), "Changes in erythrocyte sedimentation rate, CRP and hematological parameters in patients with acute malaria". Scand J Infect Dis 21(4): 4334-41].

[0084] It may be worthwhile to note, though it is not essential to the working of the present invention, that in exemplary neonatal sepsis work-ups, serial CRP tests on Day 1 and 2 may have been generally thought to afford a negative predictive value of about 99%, while the positive predictive value of one CRP test may be about 35% [Benitz et al. (1998), "Serial C-Reactive Protein Levels in the Diagnosis of Neonatal Infection". Pediatrics 102, e41, 1998].

[0085] Some of the prior art studies on CRP in association with paediatric SBI may be summarized in the table shown in Figure 7. Some, if not all, of the studies dealt with in the table shown in Figure 7 may also be described hereinbelow.

[0086] A recent prior art prospective study may have been conducted of about 408 Italian children aged between about 1 week and about 36 mos. The median age may have been about 10 mos. About 26% may have been of < about 3 mos. The studied group may have been a heterogeneous group of children presenting a fever without source. About 53% may have been hospitalized, and about 23% may ultimately have had SBI. PCT, CRP, WBC and/or ANC may have been compared on sensitivity and/or specificity for SBI. Only CRP and/or PCT may have been determined to be significant predictors of SBI. Cut-offs used may have been about 10,470 for WBC, about 6,450 for ANC, about 32 mg/L for CRP, and/or about 0.8 ng/L for PCT. Results for CRP may have been less discriminatory (P ~ .05) for those patients which may have had a fever evolution < about 8 hrs. PCT may not have been just a marker of infection, but may have been a slightly more accurate marker of severity of infection. PCT may have seemed to be a more accurate predictor at the beginning of an infection, whereas CRP, if it may have been used in a way which may have taken into account the additional time to rise in the bloodstream, may have been determined to be a better screening test, perhaps because of overall better sensitivity and/or feasibility (e.g., lower cost and/or better availability). Age may not have changed the sensitivity and/or specificity of CRP and/or PCT. Of note, about 90% of Italian children < about 3 yo may have been vaccinated against H flu and about 20% may have been vaccinated against S pneumoniae. [Andreola, B. (2007), "Procalcitonin and CRP as Diagnostic Markers of Severe Bacterial Infections in Febrile Infants and Children in the Emergency Department". Pediatr Infect DIs J 26(S): 672-677.]

[0087] A prior art study of about 99 French children with non-specific fever, aged between about 1 week and about 3 yrs, may have found that a CRP level of 4 about mg/dl may have been about 76% sensitive and/or about 79% specific in detecting SBI. CRP and/or procalcitonin may have performed relatively better than band count in predicting SBI. [Galetto-Lacour, A. (2003), "Bedside Procalcitonin and CRP Tests in Children with Fever without Localizing Signs of Infection Seen at a Referral Center". Pediatrics 1 12: 1054-1060.]

[0088] In another prior art study of about 124 Swiss children, aged between about 1 week and about 36 mos, with non-localizing fever, C-reactive protein may have been determined to have performed better than WBC in predicting SBI, but perhaps not as well as PCT. When both markers may have been associated (either above the cut-off), sensitivity may have been found to have increased to about 96% (between about 82% and about 100%), but specificity may have been found to have dropped to about 67% (between about 56% and about 76%). PPV may have been determined to have risen to about 98%, and NPV may have been determined to have dropped to about 46%. [Gendrel 2001.]

[0089] A prior art prospective study of CRP as a distinguisher of viral illness as opposed to occult bacterial illness in febrile children, aged between about 3 months and about 36 months, without apparent focus, may have found that CRP of > about 4.4 mg/L may have had about 63% sensitivity and about 86% specificity. ANC may have been determined to have better predictive value for occult bacteremia than CRP. The purportedly marginal benefit of CRP over ANC and/or WBC may have been determined to have been very limited and/or to not add substantial accuracy to either of these tests. Only about 3 children in the study may have had occult bacteremia. [Isaacman (2002), "Utility of the Serum CRP for Detection of Occult Bacterial Infection in Children". Arch Pediatr Adolesc Med 156: 905-90.]

[0090] In a prior art prospective cohort study of about 100 Indian children, aged between about 3 mos and about 36 mos, with fevers > about 38.5 ⁰C without focus (with perhaps about 25% ultimately having been bacteremic), CRP > about 4 mg/dl was about 95% sensitive and about 86% specific for the diagnosis of bacteremia. None of the other acute phase reactants individually, and/or in combination, may have been determined to have better sensitivity and/or specificity than CRP. [Kohli, V. et al. (1993), "Value of serum CRP concentrations in febrile children without apparent focus". Ann Trop Paediatr 13(4): 373-8.]

[0091] One prior art Israeli study which may have used the QuickRead test to predict bacterial gastroenteritis in about 44 children may have found that a CRP cut-off value of about 95 mg/L may have had a sensitivity of about 87%, and a specificity of about 92% for predicting culture-confirmed bacterial gastroenteritis. An RC)C result may have been about .943. Mean CRP levels of patients with bacterial gastroenteritis may have been about 224 mg/L, and that of patients with viral gastroenteritis may have been about 30 mg/L (p < about 0.001), and the LHR may have been about 1.49 (from between about 1.13 and about 1.96). PPV may have been about 70%, and/or NPV may have been about 97%. |Marcus, N. et al. (2007), "The QuickRead CRP Test ibr the Prediction of Bacterial Gastroenteritis in lhc Pediatric Emergency Department". Pediatric Emerg Care 23(9): 634-637.]

[0092] Previously, in about 119 American children aged between about 1 mo and about 36 mos, CRP may have been determined to have performed better than WBC > about 15K, and/or ANC > about 10K₅ in predicting severe bacterial illness (SBI). All three biomarkers may have been found to be more predictive ol^' SBl, ii^" a duration of a fever may have been > about 12 hrs, but WBC and/or ANC may not have been found to be reliable indicators of SBI even at fever durations of > about 12 hrs, and/or CRJ¹ may have been found to perform better regardless of a length of the fever. [Pratt, A. et al. (2007), "Duration of fever and markers of serious bacterial infection in young febrile children". PeJs Intl 49: 31-35.]

[0093] In contrast, another prior art prospective study of about 77 febrile children, aged between about 1 month and about 36 months, may have found CRP > about 7 mg/L to have a 79% sensitivity and/or a 91% specificity. CRP > about 9 rag/L may have been found to have an odds ratio of about 9. CRP may have been found to have a better predictive value than ANC and/or WBC. A relatively high prevalence of SBI might have been found to denote a selection bias. [Pulliam, P. et al. (2001), "CRP in febrile children 1 to 36 months of age with clinically undetectable ssseious bacterial infection". Pediatrics 108: 1275-1279.] [0094] CRP may have been previously found to not accurately discriminate between bacterial and viral infections in children with meningitis [e.g., Clark, D. et al. (1983), "Use of serum C-reactive protein in differentiating septic from aseptic meningitis in children". J Pediatr 1983, 102: 718-720; Hansson, L.O. et al. (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25: 625-630; and/or Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric InfDis J 18: 875-881], particularly if the CRP level may have been between about 10 mg/L and about 50 mg/L [Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric InfDis J IS: 875-881].

[0095] The prior art concerning adults may have provided reasonable support for the use of CRP in the diagnosis of sepsis, and/or for the use of serial CRP tests in judging severity of inflammation and/or outcome prediction. The table shown in Figure 8 [which may have been reproduced, in whole or in part, from Povoa, P. (2002), "CRP: a valuable marker of sepsis". Int Care Med DOI 10.1007/sOOl 34-002-1209-6 (online journal)] may summarize numerous C- reactive protein (CRP) cut-offs of different infectious situations, sensitivity and specificity.

Procalcitonin (PCT)

[0096] Procalcitonin (PCT) may be a biomarker of host response to inflammation and/or infection. Although its specific function may be unknown, PCT may help to sustain inflammatory reactions after its production in hepatocytes may have been stimulated by other acute phase reactants, such as by a tumor necrosis factor. [Assicot (1993), "High serum procalcitonin concentrations inpatients with sepsis and infection". Lancet 341(8844): 515-8.] Though not essential to the working of the present invention, it may be generally believed that procalcitonin may be about 116 amino acids in length. It may be cleaved into calcitonin by specific enzymes, which may be present in the C cells of the thyroid. Procalcitonin may be secreted about 3 hours after an intravenous injection of endotoxin. Its concentration may reach a plateau in about 6 hours and may remain high for at least about 24 hours [Dandonna et al (1994), "PCT increases after endotoxin injection in normal subjects". J Clin Endocrin Metab 79: 1605-1608]. It may be very stable, it may have a relatively long half-life (about 20 hours), and/or it may be simple to determine by the antibody sandwich method. Normal values in European patients may be < about 0.5 ng/mL [Chiwikata, C. et al. (2001), "PCT as a parameter of disease severity and risk of mortality in patients with P. falciparum malaria". J Inf Dis 183: 1161-1164]. PCT may rise slightly in viral infections and/or inflammatory conditions, but it may increase nearly about 1000-fold in bacterial infections. In infections, PCT levels may rise earlier than CRP levels. Administration of antibiotics may appear to decrease PCT levels. Calcitonin and/or its prohormone, procalcitonin, may be high in cases of thyroid medullary carcinoma and/or in some pulmonary cancers.

Prior Art uses of PCT in Malaria Endemic Areas

[0097] Elevated procalcitonin may have been previously observed in cases of malaria which may have been caused by P. falciparum. The elevated PCT may have been mediated by the production of relatively large amounts of tumor necrosis factor. [See, for example, Grau, G. et al. (1989), "Tumor necrosis factor and disease severity in children with falciparum malaria". NEJM 1586-1591; and Richard-Lenoble 1997.] [0098] There may have been a few prior art studies examining procalcitonin (PCT) as a marker and/or predictor of the severity of malaria. A study of 66 German patients with P. falciparum may have found serum PCT concentrations to be highly correlated with the absence of semi-immunity, parasite burden, the degree of disease severity, and/or mortality. PCT concentrations of > about 25 ng/mL may have been found to be highly correlated with a risk of death. [Chiwakata 2000.]

[0099] Median PCT levels in about 27 Thai patients, with complicated malaria on admission, may have been previously found to be about 40 ng/ml (0.04-662). With treatment, the median may have been found to fall to about 1.3 (between about 0.01 and about 6.5). PCT levels may have been previously found to be correlated with initial parasite density (p < about .05). [Hollenstein, U. et al. (1998), "Serum procalcitonin levels in severe P. falciparum malaria". Am J Trop Med 59(6): 860-3.]

[0100] However, for P vivax, the correlation between PCT levels and disease severity may have been previously determined to be non-existent. [Manegold et al. (2003), "PCT levels in tertian malaria". Malar J 16, 2: 34. Epub 2003 Oct 16.]

[0101] In a French study of about 25 adults, with uncomplicated P.falciparum malaria, PCT may have been found to be often (but not universally) elevated, especially when there may have been a delay between first symptoms and diagnosis, and/or when parasitemia may have been high. [Uzzan 2005.] PCT as Predictor of Severe Bacterial Infection in the Prior Art

[0102] Since about the time it may have been originally described in children [Assicot (1993), "High serum procalcitonin concentrations inpatients with sepsis and infection". Lancet 341(8844): 515-8], several papers may have demonstrated the usefulness of procalcitonin as a potential indicator of bacterial infection. In most prior art studies, procalcitonin may have been found to be at least about as good as CRP, and often better than CRP, when it may come to accuracy in predicting SBI, infection severity, and/or presence of SBI when fever duration is < about 12 hrs [e.g., Dubos, F. (2006), "Serum PCT and other biologic markers to distinguish between bacterial and aseptic meningitis". J Pediatr 149(72-6); Hansson, L. O. et al. (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25: 625- 630; Hsiao, A. et al. (2005), "Fever in the new millennium: a review of recent studies of markers of serious bacterial infection in febrile children". Curr Opin Peds 17: 56-61 ; Galetto-Lacour, A. (2001), "Precalcitonin, IL 6, IL-8, IL-I receptor antagonist and CRP as identificators of serious bacterial infections in children with fever without localising signs". Eur J Pediatr 160: 95-100; and/or Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric Inf Dis J 18: 875-881]. Usually, procalcitonin may also have been found to be better than CRP in differentiating between bacterial and viral infections [e.g., Clarke, D. et al. (1983), "Use of serum C-reactive protein in differentiating septic from aseptic meningitis in children". J Pediatr 1983; 102:718- 720; Dubos, F. (2006), "Serum PCT and other biologic markers to distinguish between bacterial and aseptic meningitis". J Pediatr 149(72-6); and/or Ip, M. et al. (2007), "Value of serum procalcitonin, neopterin, and CRP in differentiating bacterial from viral etiologies in patients presenting with Io were respiratory tract infections". Diagn Microbiol & Inf Dis 59: 131-136]. [0103] Procalcitonin concentrations may have been previously found to be high in cases of early and/or late neonatal bacterial infections [e.g., Gendrel 1996, and/or Chiesa 1998], and may sometimes have been used with CRP in North American hospitals in neonatal sepsis workups, although its expense may have limited its use [Polen, R. (2005), "Neonatal Sepsis". NICUniversity On-line CME Course].

[0104] Recent studies may have shown that, when used in conjunction with other clinical information, PCT levels may help clinicians distinguish bacterial from non-bacterial infections, and preferably thereby, decrease the inappropriate and/or unnecessary use of antibiotics dramatically, preferably without significant increased risk to the patient. [Briel, M. et al. (2008) "Procalcitonin-Guided Antibiotic Use vs a Standard Approach for Acute Respiratory Tract Infections in Primary Care". Arch Intern Med 168(18): 1-8; Simon, L. et al. (2008) "Procalcitonin and C-reactivc protein as markers of bacterial infection in critically ill children at onset of systemic inflammatory response syndrome". Pediatr Crit Care Med 9(4): 407-413.]

[0105] Studies of PCT as a predictor of SBI in children, may be summarized in the table shown in Figure 9, with cut-offs. Selected studies in the table shown in Figure 9 may also be summarized below and/or elsewhere herein.

[0106] In a prior art study of about 700 Swiss children, aged between about 1 week and about 36 mos, with nonlocalizmg fever, a PCT cut-off value of about 1 pg/L may have been found to be the best compromise between sensitivity (> about 83%) and specificity (about 93%) for distinguishing between bacterial and viral infections. [Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric Inf Dis J 18: 875-881.] The table shown in Figure 10 may have been adapted, in whole or in part, from an article based on this prior art study.

Prior Art uses of PCT in Adults

[0107] A prior art meta-analysis of thirteen blinded studies of procalcitonin, as a diagnostic test for sepsis in critically ill non-immuno-compromised adults, may have found PCT to be a good biologic marker for sepsis, severe sepsis and/or septic shock. Such analysis may have previously found PCT to be superior to CRP, although perhaps by no means a gold standard, and/or that it may perhaps be appropriate to include PCT in diagnostic guidelines for sepsis and/or in intensive care. An odds ratio for elevated PCT levels among infected patients may have been previously determined to be about 28.6, as may be compared with about 4.5 for CRP. Among eight studies which may have compared PCT to CRP, the average odds ratios may have been previously determined to have been about 4.6, P ~ 4.1. [Uzzan, B. et al. (2006), "Serum PCT in uncomplicated falciparum malaria. A preliminary study". Trav Med & Inf Dis March 2006, pp 77-80.] PCT may have been previously found to be a good marker for septic shock, and/or to be a better indicator than CRP for differentiating cardiogenic shock from septic shock [e.g., De Werra, I. (1997), "Cytokines, nitrate. nitrate, soluble tmor necrosis factor receptors, and procalcitonin cincentrations: comparisions in patients with septic shock, cardiogenic shock and bacterial pneumonia"; and/or Benoist 1998]. In adult respiratory distress syndrome [Karzai 1997] and/or in acute pancreatitis [Grau 1997], PCT may have been previously found to distinguish between bacterial and/or toxic causes. PCT may have been found to be a good marker of bacterial infections in ICUs and/or in surgical wards, after trauma and/or in cases of febrile neutropenia [Bernard 1998]. PCT may have been previously used to diagnose bacterial infections in AIDS patients [Gerard 1997]. PCT may have been found to be a good marker for distinguishing infection from rejection in transplant medicine [Staehler 1997]. In adults with bacterial meningitis, a serum PCT level of about 0.2 mg/mL may have been found to have a sensitivity and/or specificity of close to about 100% in differentiating bacterial meningitis from viral meningitis [Viallon 1999].

Prior Art Comparisons of CRP and PCT as Predictors of Severe Bacterial Infection

[0108] In the prior art, CRP may have been found to increase later than PCT. This factor may perhaps account for several authors which may have previously advised caution with the interpretation of CRP values in fever < about 8 hrs to about 12 hrs. PCT may have previously seemed to offer a slight advantage over CRP, perhaps because of its earlier increase after stimulation, and/or its comparatively better negative predictive value. In addition, PCT may also have been considered not just as a marker of infection, but also as a marker of severity of infection. This factor may perhaps account for its comparatively better performance relative to CRP, which may have been previously reported in populations with high rates of invasive infections such as meningitis and/or sepsis. [See, for example, Carrol, E. D. et al. (2002), "Procalcitonin as a diagnostic marker of meningococcal disease in children presenting with fever and a rash". Arch Dis Child 85:282-285; Fernando-Lopez 2001 ; and/or Andreola, B. (2007), "Procalcitonin and CRP as Diagnostic Markers of Severe Bacterial Infections in Febrile Infants and Children in the Emergency Department". Pediatr Infect DIs J 26(8): 672-677.]

[0109] In a prior art study of febrile adults with and without documented bacterial and/or fungal infection, a PCT level of > about 1.2 ng/mL may always have been considered as evidence of bacterial infection, and/or may have been held to be more discriminative than WBC and/or CRP, in distinguishing bacterial from other inflammatory causes [Delavaux 2003]. Prior Art Combinations of CRP and PCT to predict Severe Bacterial Infection

[0110] It may have been previously considered that use of both CRP and PCT may help to increase discriminatory power with regard to differentiating bacterial infection from viral infection [e.g., Ip, M. et al. (2007), "Value of serum procalcitonin, neopterin, and CRP in differentiating bacterial from viral etiologies in patients presenting with lowere respiratory tract infections". Diagn Microbiol & InfDis 59: 131-136; and/or Polen, R. (2005), "Neonatal Sepsis". NICUniversity On-line CME Course].

[0111] Previously, in adult patients undergoing cardiac surgery, PCT may have appeared to be useful in discriminating between acute phase response following surgery and systemic postoperative infection. Additional CRP may have been previously held to additionally increase the specificity. A high PCT level combined with a high CRP level may have been previously held to predict systemic infection. A low PCT combined with a high CRP may have previously held to predict acute phase response and/or local wound infection, but perhaps not a systemic infection. [Rothenburger, M. et al. (1999), "Detection of Acute Phase Response and Infection. The Role of Procalcitonin and CRP". Clin Chem and Lab Med 37(3): 275-279.]

Angiopoietin-2 in Malaria

[0112] Angiopoietin-2 (Ang-2) is an endothelium-specific growth factor that destabilizes vascular endothelium and increases vascular leakage, in so doing contributing to the pathophysiology of severe and cerebral malaria. Angiopoietin 2 levels may be increased in the serum of both adult and pediatric patients with cerebral malaria compared to that of uncomplicated malaria and healthy controls. ROC curves may indicate that Angiopoietin-2 levels discriminate between uncomplicated and cerebral malaria. Angiopoietin-2 levels may help to predict survival in African children with cerebral malaria. [Lovegrove, F. et al. "Serum angiopoietin-1 and -2 levels discriminate cerebral malaria from uncomplicated malaria and predict clinical outcome in African children". PLoS (in press).] Plasma Angiopoietin-2 levels may also be associated with poor clinical outcaome in severe falciparum malaria. [Yeo, T. et al. (2008) "Angiopoietin 2 is associated with decreased endothelial nitric oxide and poor clinical outcome in severe falciparum malaria". PNAS 105(44): 17097-17102.] Figures HA and HB, taken together, show a table summarizing selected prior art studies on Ang-1, Ang-2 and Ang- 2/Ang-l ratio levels in association with malaria and sepsis; and Figure 12 shows a table summarizing selected prior art studies on Ang-1, Ang-2 and Ang-2/Ang-l ratio levels in association with malaria.

Angiopoietin-2 in Sepsis and Septic Shock

[0113] In children, plasma Ang-2 levels may be significantly elevated in septic shock when compared to healthy children and critically ill children with either SIRS or sepsis, and may correlate with disease severity and outcome [Giuliano, J.S. Jr, et al. (2007) "Admission angiopoietin levels in children with septic shock". Shock 28(6): 650-654]. In adult ICU patients, Angiopoietin-2 may strongly predict mortality [Kumpers, P. et al. (2008) "Excess circulating angiopoietin 2 is a strong predictor of morality in critically ill medical patients". Crit Care 12(6): R 147 Epub; Siner, J. et al. (2008) "Increased serum angiopoietin 2 levels are associated with increased mortality in sepsis" Shock Sep 1 1, Epub ahead of print], and in septic ICU patients, Angiopoietin-2 may correlate with disease severity [Orfanos, S. et al. (2007) "Angiopoietin 2 is increased in severe sepsis: correlation with inflammatory mediators". Crit Care Med 35(1): 199- 206]. Background Conclusion

[0114] One of the objectives of an aspect of the present invention is to provide a malaria panel diagnostic device, system and method.

[0115] More generally, one of the main objectives of an aspect of the present invention is to provide a system and/or method to generate diagnostic profiles.

[0116] An objective of one aspect of the present invention may be to provide for the rapid detection of P .falciparum and/or non-falciparum malaria infections in pediatric and/or adult patients with acute febrile illness.

[0117] There may be a need for a rapid point-of-care CRP device in North America and/or around the world. Physicians having ordinary skill in the art may perceive a need for a rapid test for CRP with or without malaria.

[0118] It is an object of the present invention to obviate or mitigate one or more of the aforementioned mentioned disadvantages associated with the prior art, and/or to achieve one or more of the aforementioned objects of the invention.

SUMMARY OF THE INVENTION

[0119] According to the invention, there is disclosed a rapid diagnostic test panel for use in the diagnosis of malaria and severe bacterial infection in a test sample. The panel includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P .falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2; and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin.

[0120] According to an aspect of one preferred embodiment of the invention, P. falciparum aldolase may preferably be the first group member, Angiopoietin-2 may preferably be the second group member, and/or procalcitonin may preferably be the third group member.

[0121] According to the invention, there is also disclosed a kit for use in the diagnosis of malaria and severe bacterial infection in a test sample. The kit includes, in package form, monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P .falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin.

[0122] According to an aspect of one preferred embodiment of the invention, P. falciparum aldolase may preferably be the first group member, Angiopoietin-2 may preferably be the second group member, and/or procalcitonin may preferably be the third group member.

[0123] According to the invention, there is also disclosed a rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample. The device includes a receiving portion adapted to receive a panel which includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P. falciparum aldolase and pan P .falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin. The device also includes a sample chamber adapted to receive the sample for contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member. The device also includes a detection element for detecting signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member. The device also includes one or more processors operatively encoded with an algorithm for analysis of the signals to, on the basis of the reactions detected, automatically generate a test result which differentiates between (i) P. falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P .falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria, (iii) non-P. falciparum malaria, (iv) infection which is likely to be said severe bacterial infection, and (v) infection which is unlikely to be said severe bacterial infection, in the sample.

[0124] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. Preferably, when the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or above a predetermined second group high cut-off level, the processors may preferably generate the test result as indicative of the aforesaid P.falciparum malaria which is likely to be at risk for the aforesaid severe and/or cerebral malaria.

[0125] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. Preferably, when the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or below a predetermined second group low cut-off level, the processors may preferably generate the test result as indicative of the aforesaid P .falciparum malaria which is unlikely to be at risk for the aforesaid severe and/or cerebral malaria.

[0126] According to an aspect of one preferred embodiment of the invention, preferably when the reactions are detected with the monoclonal antibodies to the first group member but preferably not with the monoclonal antibodies to HRP2, the processors may preferably generate the test result as indicative of the aforesaid non-P. falciparum malaria.

[0127] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. Preferably, when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified at and/or above a predetermined third group high cut-off level, the processors may preferably generate the test result as indicative of the aforesaid infection which is likely to be the aforesaid severe bacterial infection.

[0128] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. Preferably, when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified at and/or below a predetermined third group low cut-off level, the processors may preferably generate the test result as indicative of the aforesaid infection which is unlikely to be the aforesaid severe bacterial infection.

[0129] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to — preferably when the processors generate the test result as indicative of the aforesaid infection which is unlikely to be the aforesaid severe bacterial infection - also generate the test result as indicative of a localized bacterial infection and/or a viral infection.

[0130] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, also include a presentation element adapted for presentation to a user of treatment and/or follow-up information related to the test result.

[0131] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. The device may preferably, but need not necessarily, also include a presentation element adapted for presentation to a user of treatment and/or follow-up information - preferably, when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified between a predetermined second group high cut-off level and a predetermined second group low cut-off level. [0132] According to an aspect of one preferred embodiment of the invention, the algorithm may preferably, but need not necessarily, operatively encode the processors to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. The device may preferably, but need not necessarily, also include a presentation element adapted for presentation to a user of treatment and/or follow-up information - preferably, when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified between a predetermined third group high cut-off level and a predetermined third group low cut-off level.

[0133] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, be adapted for use with P .falciparum aldolase as the first group member, Angiopoietin-2 as the second group member, and/or procalcitonin as the third group member.

[0134] According to an aspect of one preferred embodiment of the invention, the second group high cut-off level, the second group low cut-off level, the third group high cut-off level, and/or the third group low cut-off level may preferably, but need not necessarily, be predetermined based on local normative data.

[0135] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group high cut-off level may preferably, but need not necessarily, be about 2.0 nanograms per millilitre (ng/mL). [0136] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group low cut-off level may preferably, but need not necessarily, be about 0.5 nanograms per millilitre (ng/mL).

[0137] According to an aspect of one preferred embodiment of the invention, the sample chamber may preferably, but need not necessarily, be adapted to contact the sample with the monoclonal antibodies inside the sample chamber, and/or to flow the sample into contact with the monoclonal antibodies outside of the sample chamber.

[0138] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, be a handheld portable rapid diagnostic test device, which may preferably be adapted for use at a point-of-care.

[0139] According to an aspect of one preferred embodiment of the invention, the signals may preferably, but need not necessarily, be light signals. The detection element may preferably, but need not necessarily, be an optical detection element.

[0140] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, also include an onboard receiver element adapted to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database. The algorithm may preferably, but need not necessarily, automatically generate the test result at least in part on the basis of the clinical, epidemiological and/or sociological data. [0141] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, also include an onboard transmission element adapted to remotely transmit the test result, preferably for storage in a remote database.

[0142] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, also include an onboard geolocalization element adapted to generate a geographic location result, preferably based on the geographic location of the device. The device may preferably, but need not necessarily, also include an onboard transmission element adapted to remotely transmit the test result, preferably together with the geographic location result, and preferably for storage in a remote database.

[0143] According to an aspect of one preferred embodiment of the invention, the device may preferably, but need not necessarily, be adapted for use with whole blood, plasma and/or serum as the sample.

[0144] According to the invention, there is also disclosed a method of operating a rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample. The method includes the step of loading a receiving portion of the test device with a panel which includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P. falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin. The method also includes the step of loading the sample into a sample chamber of the test device. The method also includes the step of using the test device to contact the sample with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member. The method includes the step of using a detection element of the test device to detect signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member. The method also includes the step of using one or more processors onboard the rapid diagnostic test device to analyze the signals and automatically differentiate between (i) P .falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P .falciparum malaria which is unlikely to be at risk for severe or cerebral malaria, (iii) non-P. falciparum malaria, (iv) infection which is likely to be a severe bacterial infection, and (v) infection which is unlikely to be a severe bacterial infection, in the sample.

[0145] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. Preferably, when the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or above a predetermined second group high cut-off level, the processors may preferably be used to generate the test result as indicative of the aforesaid P. falciparum malaria which is likely to be at risk for the aforesaid severe and/or cerebral malaria.

[0146] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. Preferably, when the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or below a predetermined second group low cut-off level, the processors may preferably be used to generate the test result as indicative of the aforesaid P .falciparum malaria which is unlikely to be at risk for the aforesaid severe and/or cerebral malaria.

[0147] According to an aspect of one preferred embodiment of the invention, preferably when the reactions are detected with the monoclonal antibodies to the first group member but preferably not with the monoclonal antibodies to HRP2, the processors may preferably be used to generate the test result as indicative of the aforesaid non-P. falciparum malaria.

[0148] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. Preferably, when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified at and/or above a predetermined third group high cut-off level, the processors may preferably be used to generate the test result as indicative of said infection which is likely to be the aforesaid severe bacterial infection.

[0149] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. Preferably, when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified at and/or below a predetermined third group low cut-off level, the processors may preferably be used to generate the test result as indicative of the aforesaid infection which is unlikely to be the aforesaid severe bacterial infection.

[0150] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also includes a step of using the processors to, preferably when said infection is differentiated as unlikely to be the aforesaid severe bacterial infection, automatically characterize the sample as indicating localized bacterial and/or a viral infection.

[0151] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also include a step of using a presentation element onboard the test device to present treatment and/or follow-up information related to the test result to a user.

[0152] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. The method may preferably, but need not necessarily, also include a step of using a presentation element onboard the test device to present treatment and/or follow-up information to a user, preferably when the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified between a predetermined second group high cut-off level and a predetermined second group low cut-off level.

[0153] According to an aspect of one preferred embodiment of the invention, the processors may preferably, but need not necessarily, be used to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. The method may preferably, but need not necessarily, be also include a step of using a presentation element onboard the test device to present treatment and/or follow-up information to a user, preferably when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified between a predetermined third group high cut-off level and a predetermined third group low cut-off level.

[0154] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, be adapted for use with P .falciparum aldolase as the first group member, Angiopoietin-2 as the second group member, and/or procalcitonin as the third group member.

[0155] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also include one or more steps of predetermining the second group high cut-off level, the second group low cut-off level, the third group high cut-off level, and/or the third group low cut-off level based on local normative data.

[0156] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group high cut-off level may preferably, but need not necessarily, be about 2.0 nanograms per millilitre (ng/mL).

[0157] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group low cut-off level may preferably, but need not necessarily, be about 0.5 nanograms per millilitre (ng/mL).

[0158] According to an aspect of one preferred embodiment of the invention, the test device may preferably, but need not necessarily, be used to contact the sample with the monoclonal antibodies inside and/or outside of the sample chamber.

[0159] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, be adapted for use at a point-of-care, preferably with a handheld portable device as the test device.

[0160] According to an aspect of one preferred embodiment of the invention, the detection element may preferably, but need not necessarily, be used to detect light signals as the signals.

[0161] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also include a step of using a receiver element onboard the test device to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database. The processors may preferably, but need not necessarily, be used to generate the test result at least in part on the basis of the clinical, epidemiological and/or sociological data.

[0162] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also include a step of using a transmission element onboard the test device to remotely transmit the test result, preferably for storage in a remote database. [0163] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, also includes a step of using a geolocalization element onboard the test device, preferably to generate a geographic location result based on the geographic location of the device. The method may preferably, but need not necessarily, also include a step of using a transmission element onboard the test device to remotely transmit the test result, preferably together with the geographic location result, and preferably for storage in a remote database.

[0164] According to an aspect of one preferred embodiment of the invention, the method may preferably, but need not necessarily, be adapted for use with whole blood, plasma and/or serum as the sample.

[0165] According to the invention, there is also disclosed a computer readable medium on which is stored instructions which, upon execution, will operate a rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample. The instructions include instructions for contacting a panel received by the test device with the sample received within a sample chamber of the test device. The panel includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P. falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin. The instructions also include instructions for using a detection element of the test device to detect signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member. The instructions also include instructions for one or more processors onboard the test device to analyze the signals and automatically differentiate between (i) P .falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P. falciparum malaria which is unlikely to be at risk for severe or cerebral malaria, (iii) non- P.falciparum malaria, (iv) infection which is likely to be a severe bacterial infection, and (v) infection which is unlikely to be a severe bacterial infection, in the sample.

[0166] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions: to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member; and/or to generate the test result as indicative of the aforesaid P. falciparum malaria which is likely to be at risk for the aforesaid severe and/or cerebral malaria, preferably when (a) the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or (b) the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or above a predetermined second group high cut-off level.

[0167] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions: to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member; and/or to generate the test result as indicative of the aforesaid P .falciparum malaria which is unlikely to be at risk for the aforesaid severe and/or cerebral malaria, preferably when (a) the reactions are detected with the monoclonal antibodies to HRP2 and/or to the first group member, and/or (b) the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified at and/or below a predetermined second group low cut-off level. [0168] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions to generate the test result as indicative of the aforesaid non-P. falciparum malaria, preferably when the reactions are detected with the monoclonal antibodies to the first group member but preferably not with the monoclonal antibodies to HRP2.

[0169] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions: to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member; and/or to generate the test result as indicative of the aforesaid infection which is likely to be the aforesaid severe bacterial infection, preferably when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or (b) the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified at and/or above a predetermined third group high cut-off level.

[0170] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions: to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member; and/or to generate the test result as indicative of the aforesaid infection which is unlikely to be the aforesaid severe bacterial infection, preferably when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or (b) the reactions with the monoclonal antibodies to the third group member are quantified and/or semi- quantified at and/or below a predetermined third group low cut-off level. [0171] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions to automatically characterize the sample as indicating localized bacterial and/or a viral infection when the aforesaid infection is differentiated as unlikely to be the aforesaid severe bacterial infection.

[0172] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, also include instructions for using a presentation element onboard the test device, preferably to present treatment and/or follow-up information related to the test result to a user.

[0173] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the second group member. The stored instructions may preferably, but need not necessarily, also include instructions for using a presentation element onboard the test device, preferably to present treatment and/or follow-up information to a user, preferably when (a) the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and/or (b) the reactions with the monoclonal antibodies to the second group member are quantified and/or semi-quantified between a predetermined second group high cut-off level and a predetermined second group low cut-off level.

[0174] According to an aspect of one preferred embodiment of the invention, the instructions for the processors may preferably, but need not necessarily, include instructions to quantify and/or semi-quantify the reactions with the monoclonal antibodies to the third group member. The stored instructions may preferably, but need not necessarily, also include instructions for using a presentation element onboard the test device, preferably to present treatment and/or follow-up information to a user, preferably when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and/or (b) the reactions with the monoclonal antibodies to the third group member are quantified and/or semi-quantified between a predetermined third group high cut-off level and a predetermined third group low cut-off level.

[0175] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, be adapted for use with P .falciparum aldolase as the first group member, Angiopoietin-2 as the second group member, and/or procalcitonin as the third group member.

[0176] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group high cut-off level may preferably, but need not necessarily, be about 2.0 nanograms per millilitre (ng/mL).

[0177] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, be adapted for use with procalcitonin as the third group member. The third group low cut-off level may preferably, but need not necessarily, be about 0.5 nanograms per millilitre (ng/mL).

[0178] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, also include instructions for using a receiver element onboard the test device, preferably to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database. The instructions for the processors may preferably, but need not necessarily, include instructions to generate the test result, preferably at least in part on the basis of the clinical, epidemiological and/or sociological data.

[0179] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, also include instructions for using a transmission element onboard the test device, preferably to remotely transmit the test result, and preferably for storage in a remote database.

[0180] According to an aspect of one preferred embodiment of the invention, the stored instructions may preferably, but need not necessarily, also include instructions for using a geolocalization element onboard the test device to generate a geographic location result, preferably based on the geographic location of the device. The stored instructions may preferably, but need not necessarily, also include instructions for using a transmission element onboard the test device, preferably to remotely transmit the test result, preferably together with the geographic location result, and preferably for storage in a remote database.

[0181] Other advantages, features and characteristics of the present invention, as well as methods of operation and functions of the related elements of the panel, kit, device, method and computer readable medium, and the combination of steps, parts and economies of manufacture, will become more apparent upon consideration of the following detailed description and the appended claims with reference to the accompanying drawings, the latter of which are briefly described hereinbelow. BRIEF DESCRIPTION OF THE DRAWINGS

[0182] The novel features which are believed to be characteristic of the panel, kit, device, method and computer readable medium according to the present invention, as to their structure, organization, use, and method of operation, together with further objectives and advantages thereof, will be better understood from the following drawings in which presently preferred embodiments of the invention will now be illustrated by way of example. It is expressly understood, however, that the drawings are for the purpose of illustration and description only, and are not intended as a definition of the limits of the invention. In the accompanying drawings:

[0183] Figures IA and IB, taken together, are a flowchart of one illustrative method according to the invention;

[0184] Figures 2A and 2B, taken together, are a flowchart of another illustrative method according to the invention;

[0185] Figure 3 is a schematic representation of a test device according to the present invention;

[0186] Figures 4 and 5 show tables of relationships between CRP levels and age, from a prior art study;

[0187] Figure 6 shows a table of CRP levels for P. falciparum patients, grouped by history into non-immune and semi-immune groups, from a prior study; [0188] Figure 7 shows a table summarizing selected prior art studies on CRP in association with paediatric severe bacterial infection;

[0189] Figure 8 shows a table summarizing CRP cut-offs for varying infectious situations, sensitivities and specificities from a prior art study;

[0190] Figure 9 shows a table summarizing selected prior art studies on PCT as a predictor of severe bacterial infection in children;

[0191] Figure 10 shows a table summarizing PCT cut-offs, sensitivities and specificities for distinguishing between bacterial and viral infections, from a prior art study;

[0192] Figures HA and HB, taken together, show a table summarizing selected prior art studies on Ang-1, Ang-2 and Ang-2/Ang-l ratio levels in association with malaria and sepsis; and

[0193] Figure 12 shows a table summarizing selected prior art studies on Ang-1, Ang-2 and Ang-2/Ang-l ratio levels in association with malaria.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0194] Preferably, a rapid diagnostic test panel and a kit are for use in the diagnosis of malaria and severe bacterial infection in a test sample (not shown). The panel preferably includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) P .falciparum aldolase and/or pan P .falciparum lactate dehydrogenase, (c) Angiopoietin-1 and/or Angiopoietin-2, and (d) C-reactive protein and/or procalcitonin. The kit includes preferably includes these reagents in package form. According to one preferred embodiment of the invention, monoclonal antibodies to P .falciparum aldolase, Angiopoietin-2, and procalcitonin are used.

[0195] Figure 3 schematically illustrates a rapid diagnostic test device 100 according to the invention, which is for use in the diagnosis of malaria and severe bacterial infection in the test sample. Preferably, the device 100 is a handheld portable rapid diagnostic test device which is suitable for use at a variety of remote points-of-care. The device 100 includes a portion 320 to receive the panel, and a chamber 310 to receive the sample for contact with the monoclonal antibodies in a reacting portion 300. The reacting portion 300 may designed such that the reagents react with the sample inside or outside of the sample chamber 310. The sample chamber 310 may be likewise adapted to flow the sample into contact with the monoclonal antibodies outside of the sample chamber 310.

[0196] The device 100 also includes a detection element 208, 212 for detecting signals from the sample in contact with the monoclonal antibodies. Preferably, the signals are light signals, and the detection element 208, 212 is an optical detection element.

[0197] The device 100 also includes a processor 108 operatively encoded with an algorithm 214 for analysis of the signals. On the basis of the reactions detected, the processor 108 automatically generates test results. The test results differentiate between (i) P .falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P.falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria, (iii) non-P. falciparum malaria, (iv) infection which is likely to be said severe bacterial infection, and (v) infection which is unlikely to be said severe bacterial infection, in the sample. [0198] The algorithm 214 preferably encodes the processor 108 to semi-quantify: (1) the reactions with the monoclonal antibodies to Angiopoietin-2 (e.g., at or above a predetermined second group high cut-off level, at or below a predetermined second group low cut-off level, or in between the two); and (2) the reactions with the monoclonal antibodies to procalcitonin (e.g., at or above a predetermined third group high cut-off level, at or below a predetermined third group low cut-off level, or in between the two). Preferably, the second group high cut-off level, the second group low cut-off level, the third group high cut-off level, and the third group low cut-off level are predetermined based on local normative data. For example, when adapted for use with procalcitonin as the third group member, the third group high cut-off level may be predetermined at about 2.0 nanograms per millilitre (ng/mL), and the third group low cut-off level may be predetermined at about 0.5 nanograms per millilitre (ng/mL).

[0199] As seen in Figures IA and 2 A, when reactions 410 are detected with the monoclonal antibodies to HRP2 and to P.falciparum aldolase, and the reactions 420 with the monoclonal antibodies to Angiopoietin-2 are semi-quantified at and/or above the second group high cut-off level, the processor 108 generates the test result as indicative of the aforesaid P.falciparum malaria which is likely to be at risk for the aforesaid severe and/or cerebral malaria. When reactions 410 are detected with the monoclonal antibodies to HRP2 and to P.falciparum aldolase, and the reactions 420 with the monoclonal antibodies to Angiopoietin-2 are semi-quantified at and/or below the second group low cut-off level, the processor 108 generates the test result as indicative of the aforesaid P.falciparum malaria which is unlikely to be at risk for the aforesaid severe or cerebral malaria. When reactions 410 are detected with the monoclonal antibodies to P.falciparum aldolase but not with the monoclonal antibodies to HRP2, the processors generate the test result as indicative of the aforesaid non-P. falciparum malaria. [0200] As shown in Figures IB and 2B, when reactions 410 are detected with neither the monoclonal antibodies to HRP2 nor to P .falciparum aldolase, and the reactions 440 with the monoclonal antibodies to procalcitonin are semi-quantified at and/or above the third group high cut-off level, the processor 108 generates the test result as indicative of the aforesaid infection which is likely to be the aforesaid severe bacterial infection. When reactions 410 are detected with neither the monoclonal antibodies to HRP2 nor to P. falciparum aldolase, and the reactions 440 with the monoclonal antibodies to procalcitonin are semi-quantified at and/or below the third group low cut-off level, the processor 108 generates the test result as indicative of the aforesaid infection which is unlikely to be the aforesaid severe bacterial infection. In this last situation, the processor 108 may also preferably generate the test result as indicative of a localized bacterial infection and/or a viral infection.

[0201] As shown in Figure 3, the device 100 preferably also includes a presentation element 110 for presenting, to a user (not shown), treatment and/or follow-up information 430 related to the test result (as indicated, generally, in Figures IA to 2B).

[0202] As shown in Figures IA to 2B, the treatment and/or follow-up information 430 may also be presented to the user: (1) when reactions 410 are detected with the monoclonal antibodies to HRP2 and to P .falciparum aldolase, and the reactions 420 with the monoclonal antibodies to Angiopoietin-2 are semi-quantified between the second group high and low cut-off levels (as shown in Figures IA and 2A); and (2) when reactions 410 are detected with neither the monoclonal antibodies to HRP2 nor to P.falciparum aldolase, and the reactions with the monoclonal antibodies to procalcitonin are semi-quantified between the third group high and low cut-off levels (as shown in Figures IB and 2B). [0203] Preferably, the device 100 includes an onboard receiver and transmission element (alternately, a "communication subsystem") 114. The communication subsystem 114 may remotely receive clinical, epidemiological and sociological data regarding the sample from a remote database (not shown). The processor 108 may then generate the test results partially on the basis of the clinical, epidemiological and sociological data.

[0204] Additionally, the test device 100 also preferably includes an onboard geolocalization element 124. The geolocalization element 124 preferably generates a geographic location result based on the geographic location of the device 100. The communication subsystem 114 may remotely transmit the test results, preferably together with the geographic location result, for storage in the remote database.

[0205] Figures IA- IB and 2A-2B show two alternate methods of operating the rapid diagnostic test device 100 for use in the diagnosis of malaria and severe bacterial infection in the test sample. (Not shown in Figures IA to 2B, the method includes the steps of: loading the receiving portion 320 with the panel; loading the sample into the sample chamber 310 of the test device; using the test device 100 to contact the sample with the monoclonal antibodies; and using the detection element 208, 212 of the test device 100 to detect signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies.) Notably, the method includes the step of using the processor 108 to analyze the signals and, in the manner aforesaid, automatically differentiate between (i) P .falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P .falciparum malaria which is unlikely to be at risk for severe or cerebral malaria, (iii) non-P. falciparum malaria, (iv) infection which is likely to be a severe bacterial infection, and (v) infection which is unlikely to be a severe bacterial infection, in the sample. [0206] The method preferably also include a step of using the presentation element 110 onboard the test device 100 to present treatment and/or follow-up information 430 to the user.

[0207] Other steps which are part of the method may also include a step of using the communication subsystem 114: (1) to remotely receive clinical, epidemiological and sociological data regarding the sample from the remote database; (2) to remotely transmit the test results for storage in the remote database; and/or (3) to remotely transmit the geographic location result for storage in the remote database.

[0208] Also within the scope of the invention is a computer readable medium (not shown) on which is stored instructions. Upon execution, the stored instructions will operate the test device 100 for use in the diagnosis of malaria and severe bacterial infection in the test sample. The instructions include: instructions for contacting the panel with the sample; instructions for using the detection element 208, 212 to detect signals from the sample in contact with the monoclonal antibodies; and instructions for the processor 108 to analyze the signals in the aforesaid manner. The instructions for the processors preferably include instructions to semi-quantify: the reactions 420 with the monoclonal antibodies to P .falciparum aldolase (as may be best appreciated from a consideration of Figures IA and 2A); and the reactions 440 with the monoclonal antibodies to procalcitonin (as may be best appreciated from a consideration of Figures IB and 2B).

[0209] The stored instructions may preferably also include instructions for using: the presentation element 110 onboard the test device 100 to present the treatment and/or follow-up information 430 to the user; and the communication subsystem 114 to receive and transmit data (e.g., including the test results and the geographic location result, among other things) in the aforesaid manner. [0210] The present invention preferably includes a system and a method to generate diagnostic profiles based on the use of the diagnostic device 100. The system and method preferably enable substantially contemporaneous identification of at least two biologic markers in a biologic sample, preferably by using the same measurement system at the same time. The measurement system may preferably provide a visual reading of the test and a method to generate diagnostic profiles. Preferably the measurement system may utilize an optical sensor 208 (as best seen in Figure 3).

[0211] Preferably, the system may also be able to correlate results of the analysis with economic, healthcare, geographic and statistics data, accessible via software, to generate one or more diagnostic profiles including clinical data, and epidemiologic and geographic information.

[0212] Other features preferably include the combination of a substantially contemporaneous measurement of biomarkers, with the geo-localization of the device 100 performing the analysis, and the use of a knowledge database to extract clinical data, epidemiologic data, and sociologic information regarding the patient and/or the disease or condition targeted.

[0213] In particular, the present invention may be useful in simultaneously detecting and/or measuring the presence of P .falciparum malaria and/or non-falciparwn malaria, as well as the levels of C-reactive protein (CRP), procalcitonin, Angiopoietin-2 (Ang-2), Angiopoietin-1 (Ang- 1), and/or the ratio Ang-2/ Ang-1.

[0214] According to the present invention, the rapid differentiation between P. falciparum and non-P. falciparum infection is preferably also possible. [0215] According to the present invention, it is preferably also possible to help clinicians distinguish between uncomplicated malaria and severe and/or cerebral malaria. In this manner, the present invention may assist the clinician to accurately prognosticate and/or appropriately triage pediatric and/or adult patients with acute malaria. Though not essential to the working of the present invention, the distinction between uncomplicated and severe and/or cerebral malaria may preferably be made possible by measuring the level of Angiopoietin-2, which may generally rise dramatically in the face of impending severe and/or cerebral malaria. Because the Angiopoietin-2 level and the malaria result may preferably be reported substantially contemporaneously, according to the present invention, the clinician may be afforded an opportunity to substantially immediately initiate the appropriate treatment, potentially saving valuable time, improving clinical outcome, and/or saving resources. The level of Angiopoietin- 1 and/or the ratio Ang-2/Ang-l may alternatively, according to the present invention, be used to assist the clinician in making the distinction between uncomplicated malaria and complicated and/or cerebral malaria.

[0216] Preferably, the present invention may aid the clinician in distinguishing acute malaria from severe bacterial infection as a cause of acute fever in a febrile pediatric and/or adult patient, especially in the absence of localizing signs on physical exam. According to the invention, measuring the intensity of host inflammatory response may help the clinician assess the likelihood of severe bacterial illness in a febrile patient with a negative malaria result. The host inflammatory response may be characterized by a semi-quantitative measurement of CRP and/or procalcitonin. Preferably, with the CRP and/or procalcitonin level and the malaria results being reported substantially contemporaneously according to the present invention, the clinician may be better afforded an opportunity to identify a severe bacterial infection substantially immediately, and to start appropriate treatment sooner, potentially saving valuable time and/or resources, and/or improving clinical outcome.

[0217] According to the present invention, it is preferably possible to monitor the efficacy of a malaria treatment by using serial CRP and/or procalcitonin results. According to the present invention, serial CRP and/or procalcitonin results may accurately reflect a change in inflammatory response, preferably because of their relatively short half-lives. According to the present invention, and because HRP2 may remain elevated for up to three weeks after malaria treatment, it may not be preferable to use HRP2 to gauge the effectiveness of treatment.

[0218] According to the present invention, it is preferably possible to monitor the efficacy of treatment for a severe bacterial infection such as sepsis. According to the present invention, serial CRP and/or procalcitonin results may accurately reflect one or more changes in the magnitude of host inflammatory response, preferably because of their relatively short half-lives. According to the invention, the assessment of changes in the magnitude of host inflammatory response may preferably be useful in neonatal populations, pediatric populations and/or adult populations.

[0219] In addition to preferably helping the clinician identify patients with a probable severe bacterial illness such as septic shock, the present invention preferably also substantially simultaneously aids the clinician to better predict the clinical course of that illness. According to the present invention, elevated Angiopoietin-2 levels preferably help the clinician to better predict, by several hours to one day, the development of septic shock in patients with SIRS and/or sepsis. According to the present invention, elevated ratio Ang-2/Ang-l is preferably also used to identify patients with severe bacterial illness at elevated risk for mortality. According to the present invention, this identification is preferably useful in neonatal populations, pediatric populations and/or adult populations.

[0220] The present invention may also find use for surveillance and/or in epidemiologic studies of malaria in endemic areas. The combination of host biomarker levels with a positive malaria result may provide a reasonable case definition of clinical malaria, especially in endemic areas where asymptomatic parasitemia may be common. Preferably, according to the present invention, this case definition may be validated prior to adoption.

[0221] Preferably, the present invention is suitable and/or adapted for use with whole blood, plasma or serum, preferably collected either via a finger and/or heel prick, or via venipuncture.

[0222] Preferably, the present invention is suitable and/or adapted for use in substantially any and/or all health care settings worldwide, including points of care without access to clinical laboratory services. Preferably, the present invention is suitable and/or adapted for use by health care providers in civilian and/or military settings worldwide. Preferably, the present invention is suitable and/or adapted for use by any and/or all health care workers who make diagnostic decisions, e.g., physicians and nurses. Preferably, the present invention is suitable and/or adapted for use by health care workers of differing levels of training in civilian and/or military settings in developed and/or developing countries. Preferably, the present invention is especially useful at the point of care, and/or at the periphery of health care systems in the developing world. It may preferably also be used in commercial and/or industrial settings where the risk of malaria is high. Perhaps notably, the present invention may be suitable and/or adapted for use by laboratory technicians and employees, military medics, persons working in malaria surveillance, and/or those conducting epidemiologic studies. In endemic areas, this number may also include pharmacy employees and/or the general public.

[0223] Variously, according to some specific embodiments of the present invention, the malaria panel diagnostic devices, systems and/or methods may involve tests for one or more the following biomarkers: (1) quantitative tests for Angiopoietin-2, Angiopoietin-1, the ratio Ang- 2/Ang-l, PCT and/or CRP; and (2a) qualitative tests for HRP2 and aldolase; or (2b) qualitative tests for HRP2 and pan LDH.

[0224] See, for example, Figures IA- IB and 2A-2B for further information concerning the tested bio-markers for the malaria panel diagnostic devices 100 according to the present invention, and related systems and/or methods for interpreting the results generated. For purposes of example, in Figures IA- IB and 2A-2B, Ang-2 and PCT have been chosen to represent the larger group of host marker measures which may include Ang-2, ratio Ang-2/ Ang- 1 , CRP and PCT, any combination of which may preferably be used according to the present invention.

[0225] Preferably, adding the host biomarkers procalcitonin, C-reactive protein and/or Angiopoietin-2 to a point-of-care malaria diagnostic test, according to the present invention, may help to greatly extend its clinical utility, preferably by providing therapeutically useful information substantially contemporaneously with the malaria result - for example, instead of during a second round of tests hours to days later. According to the present invention, these markers may preferably help the clinician appropriately manage the patient substantially immediately, preferably at the time of the malaria result - for example, instead of later. [0226] Preferably, taken in conjunction with other clinical information including physical exam, history and/or other lab tests (if available), according to the present invention, levels of the host biomarker procalcitonin may help: (i) the clinician decide, substantially contemporaneously with a negative malaria result, whether antibiotics may be indicated; and/or (ii) with time- sensitive triage decisions, such as, for example, whether to hospitalize, preferably, for example, by reporting the likelihood of the presence of a severe bacterial infection (e.g., sepsis, pneumonia and/or meningitis) versus a localized bacterial infection (e.g., otitis media) and/or a viral syndrome.

[0227] Preferably, adding the host biomarker Angiopoietin-2 to the malaria diagnostic test, according to the present invention, may add therapeutically useful information, preferably by providing the clinician (substantially contemporaneously with the malaria result) with a measure of the likelihood that the patient may go on to have complicated and/or cerebral malaria. This information may be prognostic, preferably in that the Angiopoeitin levels may typically rise, several hours to one day, in advance of the patient's actual clinical decompensation.

[0228] Preferably, the high negative predictive values of procalcitonin and Angiopoietin-2 may also be useful, according to the invention, to help confirm a clinician's impression that the patient may be managed safely as an outpatient.

[0229] One or more of the following features are preferred features of the device, system and/or method according to the present invention. In some cases, a particular feature may be optional and/or may find application in an alternate embodiment according to the present invention, rather than being preferred or otherwise desirable. • The test is preferably read visually and/or by a sensor which is preferably an optical sensor 208 (see, for example, Figure 3).

• Preferably, it is affordable and/or cost effective to employ the test device and methods. Preferably, the cost is clearly offset by savings obtained by decreased use of inappropriate treatments.

• Preferably, the test device and methods afford (1) a sensitivity > about 95% at > about 100 parasites / uL {P. falciparum), and/or (2) a sensitivity > about 95% at > about 1000 parasites / uL (P.vivax).

• The samples used in association with the test device 100 and methods are preferably easy to obtain and/or label, and/or acceptable to young children and/or parents (e.g., a finger prick).

• The assay and/or device 100 preferably uses a small volume of fresh whole blood (e.g., with no anticoagulant and/or with a standard anticoagulant, such as EDTA, ACD), such as may be acquired, for example, by a finger prick.

• The assay and/or device 100 preferably is capable of using frozen whole blood samples collected in EDTA and/or ACD. The assay is also preferably able to use plasma and/or serum instead of, and/or in addition to, whole blood.

• The test results are preferably generated rapidly enough to enable treatment at first visit - preferably < about 30 minutes, and more preferably < about 15 minutes. (Current malaria RDTs may take in the order of about 15 min.)

• The device 100 and consumables are preferably stable during poor transport and/or storage conditions. Preferably, the device and/or consumables are heat stable (10 - 40 C), and stable in conditions of high humidity, dust and/or vibration. (Preferably, the consumables do not require refrigeration, and/or cold chain, during delivery.)

• Preferably, the method and system are simple to perform. Preferably, training requirements are minimal. Preferably, a lab technician and/or trained health care provider is not required to perform the method and/or system.

• Preferably, the method and system do not require lab equipment. Preferably, for example, the method and system do not require any external water supply, and/or centrifuge, etc. Preferably, the device 100, system and method may utilize rechargeable batteries, and a solar and/or mechanical (e.g., crank) recharge.

• Preferably, the method and system are substantially and/or entirely self-contained including internal controls.

• Preferably, the consumables have a shelf life of at least about 12 months, and more preferably, closer to about 18 months.

• Preferably, the method and system maintain standards of confidentiality and/or informed consent.

• Preferably, the results are easy to read, easy to interpret and/or provide information that may affect treatment.

• Preferably, results are readily recordable in a medical record.

• Preferably, the software used according to the present invention is flexible to meet regional needs and/or conditions.

[0230] One or more embodiments of the malaria panel diagnostic device 100, system and method according to the present invention may have a very high clinical value. In fact, some embodiments of the system and method to generate diagnostic profiles and the malaria panel diagnostic device 100, system and method according to the present invention may be crucial for malaria-endemic areas. According to one aspect of the present invention, the discriminatory power of CRP tests and/or any difficulty in defining cut-off values may be addressed by: (a) reporting semi-quantitative results and/or in providing users with flexible devices, systems and/or methods with respect to whether and/or how to use the CRP results; and/or (b) adding a second biomarker [e.g., procalcitonin (PCT) which may have a slightly higher predictive accuracy for severe bacterial infection]. Though not essential to the working of the present invention, it may be believed that the use of two biomarkers may together increase clinical usefulness and/or increase sensitivity.

[0231] Persons having ordinary skill in the art (particularly those intimately familiar with the situation in Africa) may appreciate that one or more embodiments of the malaria panel diagnostic device 100, system and method according to the present invention may find advantageous application in malaria-endemic countries. For example, it may be advantageous to utilize and/or distribute the malaria panel diagnostic device 100, system and method according to the present invention in association with one or more private sector chains of labs, such as may be found in South Africa.

[0232] It may also be appreciated that one or more embodiments of the malaria panel diagnostic device 100, system and method according to the present invention may find advantageous application in North America. For example, in paediatric contexts, it may be advantageous to use the malaria panel diagnostic device 100, system and method according to the present invention to identify serious infections, potentially in association with continuing clinical education to raise awareness over the value of host biomarkers as indicators of severe bacterial infection. Similarly, the malaria panel diagnostic device 100, system and method according to the present invention may find application in the neonatal context, and/or in the contexts of tropical and/or travel medicine.

[0233] Other potential applications may be appreciated by military personnel having ordinary skill in the art.

[0234] It may be readily possible to obtain regulatory and/or other approvals (e.g., FDA approval) for one or more embodiments of the malaria panel diagnostic device 100, system and method according to the present invention. According to one aspect of the present invention, the device 100, system and method may preferably target only one pathogen. It may be generally believed (though it is not essential to the working of the present invention) that this factor may readily afford the FDA a better opportunity to concentrate on approving the platform itself. In addition, FDA-approved comparator devices may already exist for malaria, CRP and/or procalcitonin (PCT).

[0235] According to various preferred and alternate embodiments of the invention, the test device 100 may be partially embodied by a cellular telephone, a mobile communications device (e.g., a smart phone and/or a radio-frequency chipset device), a personal digital assistant, a pocket PC, a laptop computer, a desktop computer, a navigation device, a digital audio player, a camera, a gaming device, a television, and/or a radio. According to some preferred embodiments of the invention, it may be suitable to utilize any electronic device which has the preferred display element 112, and/or a capacity to run, analyze, record and/or transmit the test results.

[0236] Now, with further reference to Figure 3, and depending on the technology used for detection, the detection element 208, 212 may preferably, and by way of a non-limiting example, be optical in nature (e.g., relying on fluorescence or colorimetry) or electrical in nature (e.g., relying on impedance effects). Preferably, many different detection technologies may be capable of use within the test device 100 (and which may be capable of modification in function, in the discoveries made, and/or in the detection field), such as, for example and among other things, one or more of the following: lateral flow strip detection technologies; nano and/or micro cytometer detection technologies; impedance sensor detection technologies; dieletrophoresis detection technologies; micro PCR detection technologies; and/or electro peptide sensor technologies.

[0237] The detection element 208, 212 may include a photodiode (e.g., a charge coupled device) 208. Preferably, the photodiode 208 may work together with a light source element 212 of the test device 100 to operatively detect test data from the sample after reaction with the reagents. Any number of different light source elements 212 might be used - for example, a laser, a tungsten lamp, a mercury lamp, a xenon lamp, a light emitting diode (LED) and/or an optical fiber light source. Additionally, other types of photodiodes 208, light detectors (e.g., photodetectors) and/or sensors may be used - e.g., a CMOS camera - may be used in place thereof and/or in addition thereto.

[0238] As shown in Figure 3, onboard memory 116, 118 may preferably be provided within the test device 100. The onboard memory 1 16, 118 may preferably be used to store test software algorithms 214 required to run the test - e.g., including the test method, the analysis process, GUI interface instructions, and any other software applications or algorithms 142, 144 associated with the test. The onboard memory 116, 118 may preferably also be capable of storing the test data for use later. The onboard memory 116, 118 is preferably also associated with the processor 108, or CPU capability, onboard the test device 100. The processor 108 uses the software algorithms 214 to control the rapid diagnostic test. [0239] The processor 108 may preferably provide enough processing capability to control the test device 100. For example, the algorithms 214 may include device management software and data analysis software. The device management software may include graphical user interface (GUI) software. According to the invention, the GUI software may preferably assist, facilitate or enable display of presentation data to a user of the test device 100. The data analysis software may include test data processing and diagnostic applications. The test data processing applications may preferably include algorithms to analyze the test data. The testing of the sample by the test device 100 may be directly initiated by the processor 108 - e.g., by instructing the user to add the sample.

[0240] The processor 108 operatively receives the test data, and applies the test data processing applications to the test data to generate highly sensitive and accurate quantitative, semi-quantitative and/or qualitative test results and/or presentation data based on the test data. In so doing, according to some preferred embodiments of the invention, the test results may be semi-quantified as high, medium, and/or low results (e.g., a low intensity of infection result). Perhaps notably, the test results which are generated according to the present invention have comparable accuracy and sensitivity with those which have been previously quantified in a laboratory or hospital setting. Advantageously, therefore and due in part to the portability inherent in the handheld test device 100, the present invention enables the generation of highly sensitive and accurate quantitative, semi-quantitative and qualitative test results outside of such laboratory and hospital settings.

[0241] It may also be worthwhile to note that the presentation data presented to the user may preferably include treatment and follow-up suggestion data 430 (as may be best appreciated from a consideration of Figures IA to 2B) based on the test results. The test device 100 is preferably adapted to generate such presentation data. The treatment and follow-up suggestion data 430 is preferably determined with reference to one or more of the algorithms 214 stored on the test device 100, or in remote and/or distributed databases and/or servers (not shown).

[0242] Preferably, the onboard memory 116, 118 of the test device 100 electronically stores the test data and one or more of the algorithms 214.

[0243] The test device preferably also has a presentation element 110. The presentation element 110 preferably includes a display element 112 which has a display capability (e.g., a display screen and/or a printer) and/or which offers a graphical user interface (or GUI). Preferably, the algorithms 214 generate the quantitative, semi-quantitative and/or qualitative test results and/or the presentation data for presentation by the test device 100 in the form of visually and/or audibly presentable data. Audibly presentable data may take the form of a verbal, musical, tonal and/or other alert sounds.

[0244] Visually presentable data may take the form of text, graphics and/or colored indicator lights. One form of visually presentable data which is contemplated according to the present invention is visually presentable textual data. Among other things, the display element 1 12 might also present graphical data which may include charts and other comparative visual representations of the quantitative test results. By way of example, and among other things, visually and/or audibly presentable data may also include descriptive and/or numerical data. Exemplary types of descriptive data may include the treatment and follow-up suggestion data and/or intensity information. Intensity data may be shown in textual and/or graphical format. Exemplary types of numerical data may include the quantitative test results. Other visually presentable data may include textual data, and/or colored indicator light data. Preferably, the display element 112 enables display of the presentation data. (In some embodiments of the invention, a printer, a speaker onboard the test device 100 or other kinds of output systems are used for visualization or presentation.) The presentation element 110 operatively presents the presentation data to the user.

[0245] As shown in Figure 3, the test device 100 is preferably a networking electronic device and is provided with a communication subsystem 114 to afford connectivity and/or communications (e.g., network connection, GSM, satellite connection, Internet) capabilities. The communication subsystem 114 networks with an external network 600 which may be a satellite network (e.g., GPS networks), a terrestrial wireless network (e.g., a cellular telephone network, a local wireless network), the Internet, and/or a laboratory and/or hospital information system network. The test device 100 may preferably be in wireless (and/or wired) communication with at least one external network 600. The communication subsystem 114 which is provided may be a cellular telephone network, an intranet connection, or a wired or wireless Internet connection.

[0246] The test device 100 may preferably also have the ability to connect quickly and easily to LIS/HIS networks via, for example, the local wireless network (e.g., a Bluetooth network) and/or a USB cable. Preferably, the test device 100 automatically transmits the test data, the presentation data (e.g., test results) and/or encrypted data for recordal in one or more remote and/or distributed databases or servers (e.g., in a LIS/HIS network). Additionally, transmission of the test data, the presentation data (e.g., the test results) and/or the encrypted data by the test device, via the communication subsystem 114 over the network 600, may be initiated directly and/or indirectly by the user by controlling a dedicated button or a context dependent programmable button or key. Preferably, the test device 100 may be provided with an electronic device memory (e.g., a flash memory) 118 which is able to record the test results related to each test. The remote and/or distributed databases and/or servers may also be used for various tests or patients and are preferably linkable with the data stored on the test device 100.

[0247] Various databases may interface with the communications subsystem, preferably including, software applications databases (e.g., clinical software applications, database software applications, download portals, quality control central databases), and various test result databases (e.g., healthcare providers database, governmental agency databases, military department databases). Notably, the databases may include, without limitation, epidemiologic databases, UN and major / international healthcare institution databases, healthcare and emergency infrastructure databases, education and economic databases, news databases, demographic databases, communication and military infrastructure databases, and weather and topographic databases. The databases may preferably serve as an additional repository for the test results (test result databases), and/or as an additional source for the test device to acquire the algorithms and/or updates to the algorithms (e.g., test processing algorithms and software applications) - i.e., from software applications databases.

[0248] Communication functions may be performed through the communication subsystem 114, which preferably acts as both a receiving element and a transmitting element.

[0249] Specific device applications 142, 144, 214 may also be stored in flash memory 1 18 and may include GPS map applications 144, among others. Those skilled in the art will appreciate that the operating system, specific device applications, or parts thereof, may be temporarily loaded into a volatile store, such as the RAM 116, for processing by the processor 108. [0250] Preferably, the algorithms 214 are operatively loaded onto the test device 100. A keypad (alternately herein a "keyboard") and/or the display element 112 of the test device may preferably be utilized in association with the test device 100 and/or the algorithms 214.

[0251] The test device 100 and the methods which are provided according to the present invention are preferably adapted for use in association with infectious disease surveillance, diagnosis and emergency medical response at the point of care in developed and developing countries, refugee camps, etc.

[0252] The test device 100 and the methods which are provided according to the present invention are preferably adapted to rapidly perform quantitative, semi-quantitative and/or qualitative diagnostic tests. It will be appreciated from the disclosures herein that the test device 100 and the methods are preferably be available for deployment and/or purchase at a reasonable price - i.e., a potentially important factor in many of the areas of the world where infectious disease surveillance may be of primary concern.

[0253] As aforesaid, the test device 100 is preferably a networking electronic device. It is preferably equipped with a global communication capability (e.g., according to a GSM and/or GPRS protocol), and with a location tracking capability via the geo-localization element (e.g., a global location tracking element and/or a GPS subsystem) 124 provided onboard the test device 100.

[0254] The test device 100 and the methods are preferably adapted to enable smart patient data management, in full compliance with all applicable interface standards for exchanging and/or transferring health data - e.g., the HL7 and/or ASTM standards. [0255] Ideally, the test device 100 may preferably allow a patient and/or healthcare provider to readily perform - preferably at their fingertips and/or in the palm of their hand - one or more diagnostic tests with substantially the same analytic capability as other substantially more unwieldy prior art high-tech diagnostic devices.

[0256] Figures IA- IB and 2A-2B show, schematically by way of overview, two preferred methods of testing the sample, for use with the reagents and the test device. The sample is collected and loaded, by the user (e.g., a patient, nurse and/or doctor), into the test device.

[0257] Thereafter, the processor 108 preferably sends a request or instruction for sensing and/or detection of the test. Test detection may, for example, involve fluorescence. Preferably, however, many different types of detection technologies may be capable of use as a detection platform inside of the test device 100. The sending of the aforesaid request or instruction by the processor 108 is preferably operative, in the sensing step, to activate at least one of sensors 208 (e.g., to open an electronic shutter in the CCD to take an image) so as to detect the test data.

[0258] In a processing step, image analysis and/or biomarker concentration calculation may preferably be performed by the processor 108.

[0259] The methods may also include a storage step, in which the test data may preferably be electronically stored using the onboard memory 116, 118 of the test device 100.

[0260] In a presentation step, the processor 108 may preferably present the test data and/or the test results for display to the user by the display element 1 12. In the processing step, one or more of the algorithms 214 may preferably have been applied to the test data and/or to the test results using the processor 108 to generate either the test results and/or the presentation data based on the test results. The presentation data so generated preferably includes the treatment and follow-up suggestion data 430 based on the test results. As aforesaid, the treatment and follow-up suggestion data 430 is preferably determined with reference to one or more of the algorithms 214 stored onboard the test device 100, or in one of the remote and/or distributed databases and/or servers. Preferably, one or more of the algorithms 214 generate the test results and/or the presentation data for presentation from the display element 112 in the form of one or more visually presentable textual data, graphical data, or colored indicator light data.

[0261] Preferably thereafter, in the presentation step, the user may preferably be provided with access to the test results, preferably via the display element 112. That is, the presentation data (e.g., the test results and treatment and follow-up suggestion data 430) are presented to the user using the presentation element 110. Preferably, in the presentation step, the presentation data are presented from the display element 112 of the presentation element 110. The processor 108 may preferably be operative, in the presentation step, to display combined test information, including the test results together with geo-localization data for the test (and patient ID data) from the display element 1 12.

[0262] According to some preferred embodiments of the invention, wireless transmission to a remote and/or distributed database and/or server takes place automatically (in accordance with a networking step). In the networking step, the test device 100 may send data to a remote and/or distributed database and/or server. According to some preferred embodiments of the invention, the remote and/or distributed database and/or server may preferably (but need not necessarily) include one or more remote laboratory and/or hospital information systems. [0263] Treatment information 430 may be provided via database(s) internal and/or external to the test device 100, and presented from the presentation element 110 of the test device 100.

[0264] It may be appreciated by persons having ordinary skill in the art that, with utilization of the test device 100 and methods according to the present invention, it may preferably be possible to greatly improve upon the limited functionality which may have been afforded by prior test devices. When all of the foregoing functionalities are combined with the relatively low total cost associated with the test device 100, it may additionally be appreciated that same may be affordable for use in developing countries.

[0265] It is noted that, to the extent possible and/or permitted by law, the teachings and/or disclosures of each of the following prior art references (which may or may not be referenced elsewhere herein) are hereby specifically incorporated by reference into the present application: Adnet, F. et al (1997), "Value of CRP in the detection of bacterial contamination at the time presentation in drug-induced aspiration pneumonia." Chest 1 12: 466-471 ; Andreola, B. (2007), "Procalcitonin and CRP as Diagnostic Markers of Severe Bacterial Infections in Febrile Infants and Children in the Emergency Department". Pediatr Infect DIs J 26(S): 672-677; Aoufi, A. et al. (2000), "Usefulness of procalcitonin for diagnosis of infection in cardiac surgical patients". Crit Care Med 28: 3171-3176; Assicot (1993), "High serum procalcitonin concentrations inpatients with sepsis and infection". Lancet 341(8844): 515-8; Baker, K. et al. (2008) "Procalcitonin assay in systemic inflammation, infection and sepsis: Clinical utility and limitations". Crit Care Med 36(3): 941-952; Benitz et al. (1998), "Serial C-Reactive Protein Levels in the Diagnosis of Neonatal Infection". Pediatrics 102, e41, 1998; Black, S. et al. (2001), "Postlicensure evaluation of the effectivenesss of seven valent pneumococcal conjugate vaccine". Pediatr Infect Dis J 2001, 20: 1105-1107; Briel, M. et al. (2008) "Procalcitonin- Guided Antibiotic Use vs a Standard Approach for Acute Respiratory Tract Infections in Primary Care". Arch Intern Med 168(18): 1-8; Carrol, A. et al. (2002), "C-Reactive Protein?" Pediatrics 110(2) 442; Carrol E.D. et al. (2002), "Procalcitonin as a diagnostic marker of meningococcal disease in children presenting with fever and a rash". Arch Dis Child 85: 282-285; Cheesbrough, J. et al. (1997), "Clinical definition for invasive Salmonella infection in African children". Pediatr Infect Dis J. 16(3): 227-83; Cheval et al. (2000), "PCT is useful in predicting the bacterial origin of an acute circulatory failure in critically ill patients". Int Care Med 26: Sl 53- 158; and/or Chiwikata, C. et al. (2001), "PCT as a parameter of disease severity and risk of mortality in patients with P. falciparum malaria". J InfDis 183: 1161-1164.

[0266] Also, to the extent possible and/or permitted by law, the teachings and/or disclosures of each of the following prior art references (which may or may not be referenced elsewhere herein) are also hereby specifically incorporated by reference into the present application: Clarke, D. et al. (1983), "Use of serum C-reactive protein in differentiating septic from aseptic meningitis in children". J Pediatr 1983, 102: 718-720; Crit Care Med 25(4): 607-13; Cunha, J. et al. (1997), "CRP: a good parameter for sepsis diagnosis 9" (abstract). Intensive Care Med 23: S61; Dahler-Ericksen, B. et al. (1997), "Evaluation of a near-patient test of CRP used in daily routine in primary healthcare by use of different plots". Clin Chem 43: 2064-2075; Dandonna et al. (1994), "PCT increases after endotoxin injection in normal subjects". J Clin Endocrin Metab 79: 1605-1608; De Werra, I. (1997), "Cytokines, nitrate.nitrate, soluble tmor necrosis factor receptors, and procalcitonin cincentrations: comparisions in patients with septic shock, cardiogenic shock and bacterial pneumonia"; Dubos, F. (2006), "Serum PCT and other biologic markers to distinguish between bacterial and aseptic meningitis". J Pediatr 149 (72-6); Ericksson, B. (1989), "Changes in erythrocyte sedimentation rate, CRP and hematological parameters in patients with acute malaria". Scand J Infect Dis 21(4): 4334-41; Esposito, S. (2005), "Evaluation of a rapid bedside test for the quantitative determination of CRP". Clin Chem and Lab Med 53(4): 438-440; and/or Fendler, W. et al. (2007) "Procalcitonin in the early diagnosis of nosocomial sepsis in preterm neonates". Jl Pediatr Child Health 44: 114-118.

[0267] To the extent possible and/or permitted by law, the teachings and/or disclosures of each of the following prior art references (which may or may not be referenced elsewhere herein) are additionally hereby specifically incorporated by reference into the present application: Fernandez-Lopez, A. et al. (2003), "Procalcitonin in pediatriac emergency departments for the early diagnosis of invasive bacterial infections in febrile infants". Pediatr Inf Dis J 22: 895-903; Ford, E. et al. (2003), "C-Reactive Protein: US reference ranges from the National Health and Nutrition Examination Survey, 1999-2000" Clin Chem, 49 (1353-1357); Galetto-Lacour, A. (2001), "Precalcitonin, IL_6, IL-8, IL-I receptor antagonist and CRP as identificators of serious bacterial infections in children with fever without localising signs". Eur J Pediatr 160: 95-100; Galetto-Lacour, A. (2003), "Bedside Procalcitonin and CRP Tests in Children with Fever without Localizing Signs of Infection Seen at a Referral Center". Pediatrics 112: 1054-1060; Gendrel, D. (1999), "Comparison of procalcitonin with CRP, interleukin 6 and interferon alpha for differentiation of bacterial vs. viral infection". Pediatric Inf Dis J 18: 875-881; Gillespie, S. et al. (1991), "Measurement of acute phase proteins for assessing severity of Plasmodium falciparum malaria". J CHn Path 44(3): 228-31; Giuliano, J.S. Jr. et al. "Admission angiopoietin levels in children with septic shock". Shock 28(6): 650-654; Gomes, M. et al. (1994), "Symptomatic identification of malaria in the home and in the primary health care clinic". Bull World Health Organ 72(3): 383-90; Graninger, W. et al. (1992), "Serum protein concentration in P.falciparum malaria". Act Trop 52: 121-128; Grau, G. et al. (1989), "Tumor necrosis factor and disease severity in children with falciparum malaria". NEJM 1586-1591; Hansson, L. O. et al. (1993), "Serum CRP in the differential diagnosis of acute meningitis". Scand J Infect Dis 1993, 25: 625-630; Hausfater, P. et al. (2002), "The Usefulness of PCT as a Marker of Systemic Infection in Emergency Department Patients: A Prospective Study". 34: 895-901; Hollenstein, U. et al. (1998), "Serum procalcitonin levels in severe P. falciparum malaria". Am J Trop Med 59(6): 860-3; Hopkins, H. et al. "Effectiveness and safety of training in fever case management and malaria RDT use at health centers in Uganda". Abstract. 57^th ASTMH meeting, December 2008; Hsiao, A. et al. (2005), "Fever in the new millennium: a review of recent studies of markers of serious bacterial infection in febrile children". Curr Opin Peds 17: 56-61; Hurt, N. (1994), "Do High Levels of CRP in Tanzanian Children Indicate Malaria Morbidity?" Clin Diag Lab Immunol, July 1994: 437-444; and/or Imrie, H. (2007), "Low Prevalence of an Acute Phase Response in Asymptomatic Children from a Malaria-Endemic Area of Papua New Guinea". AM J Trop Med Hyg 76(2): 280-284.

[0268] Further, to the extent possible and/or permitted by law, the teachings and/or disclosures of each of the following prior art references (which may or may not be referenced elsewhere herein) are also hereby specifically incorporated by reference into the present application: Ip, M. et al (2007), "Value of serum procalcitonin, neopterin, and CRP in differentiating bacterial from viral etiologies in patients presenting with lower respiratory tract infections". Diagn Microbiol & In/ Dis 59: 131-136; Isaacman (2002), "Utility of the Serum CRP for Detection of Occult Bacterial Infection in Children". Arch Pediatr Adolesc Med 156: 905-90; Ishimine, P. (2006), "Fever without source in children 0 to 36 months of age". Fed Clin NA 53(2): 167-194; Kamya, M. et al. (2007), "Effects of TMP-SMZ and insecticide-treated bednets on malaria among HIV-infected Ugandan children". AIDS, 21(15): 2059-66; Kohli, V. et al. (1993), "Value of serum CRP concentrations in febrile children without apparent focus". Ann Trop Paediatr 13(4): 373-8; Koj, A. (1985). "Catabolism and turnover of acute phase proteins", p 145-160 in A.H. Gordon and A. Koj (ed), The acute phase response to injury and infection. Elsevier, Amsterdam; Kumpers, P. et al. (2008) "Excess circulating angiopoietin 2 is a strong predictor of morality in critically ill medical patients". Crit Care 12(6) : R 147 Epub; Kupperman, N. (2002), "The Evaluation of Young Febrile Children for Occult Bacteremia". Arch Pediatr Adolesc Med 156: 855-857; Kyriacou, D. (1996), "Emergency Department Presentation and Misdiagnosis of Imported Falciparum Malaria". Annals Emerg Med, 27(6): 696-699; Lovegrove, F. et al. "Serum angiopoietin- 1 and -2 levels discriminate cerebral malaria from uncomplicated malaria and predict clinical outcome in African children", (in press); Luxemburger, C. et al. (1998), "Clinical features cannot predict a diagnosis of malaria or differentiate the infecting species in children living in an area of low transmission". Trans R Soc Trop Med Hyg. 92(1): 45-9; Malvy, D. (1992), "Laser Immunonephelometry Reference Intervals for Eight Serum Proteins in Healthy Children". Clin Chem 38(3): 394-399; Manegold et al. (2003), "PCT levels in tertian malaria". Malar J 16, 2: 34. Epub 2003 Oct 16; Marcus, N. et al. (2007), "The QuickRead CRP Test for the Prediction of Bacterial Gastroenteriitis in the Pediatric Emergency Department". Pediatric Emerg Care 23(9): 634-637; Massaga, J. (1999), "Malaria presumptive treatment in Tanzania: is it rational approach for malaria management in rural health units?" Afr J Health ScL 6(1): 22-6; Muller, B. et al. (2000), "Calcitonin precursors are reliable markers of sepsis in a medical intensive care unit". Crit Care Med 28: 977-983; Naik, P. et al. (1984), "Serum CRP levels and falciparum malaria". Trans R Soc Trop Med Hyg 78: 812-813; Nordstrand, A. et al. (2007), "Tickborne relasing fever diagnosis obscured by malaria, Togo". Emerg Infect Dis 13(1): 117-23; Orfanos, S. et al. "Angiopoietin 2 is increased in severe sepsis: correlation with inflammatory mediators". CWt Care Med 35(1): 199-206; Polen, R. (2005), "Neonatal Sepsis". NlCUniversity On-line CME Course; Povoa, P. (2002), "CRP: a valuable marker of sepsis". Int Care Med DOI 10.1007/s00134-002-1209-6 (online journal); Povoa, P. et al. (1998), "CRP as an indicator of sepsis". Intens Care Med 24: 1052- 1056; Pratt, A. et al. (2007), "Duration of fever and markers of serious bacterial infection in young febrile children". Peds Ml 49: 31-35; Pulliam, P. et al. (2001), "CRP in febrile children 1 to 36 months of age with clinically undetectable ssseious bacterial infection". Pediatrics 108: 1275-1279; Rau, B. et al. (200), "Serum amyloid A versus CRP in acute pancreatitis: clinical value of an alterative acute-phase reactant". Crit Care Med 28: 736-742; and/or Ray, S. (1995), "Clinical audit of malaria diagnosis in urban primary curative care clinics, Zimbabwe". Cent Afr JMed4l(l2): 385-91.

[0269] It is additionally noted that, to the extent possible and/or permitted by law, the teachings and/or disclosures of each of the following prior art references (which may or may not be referenced elsewhere herein) are additionally hereby specifically incorporated by reference into the present application: Reinhart, K. et al. (1998), "PCT and Its Role in the Diagnosis of Sepsis". Sepsis 2: 157-161; Reyburn, H. et al. (2004), "Overdiagnosis of malaria in patients with severe febrile illness in Tanzania: a prospective study". BMJ 329(7476): 1212; Rothenburger, M. et al. (1999), "Detection of Acute Phase Response and Infection. The Role of Procalcitonin and CRP". Clin Chem and Lab Med 37(3): 275-279; Simon, L. et al. (2008) "Procalcitonin and C-reactive protein as markers of bacterial infection in critically ill children at onset of systemic inflammatory response syndrome". Pediatr Crit Care Med 9(4): 407-413; Simon, L. et al. (2004) "Serum Procalcitonin and C-Reactive Protein Levels as Markers of Bacterial Infections: A Systematic Review and Meta-analysis". Clin Inf Dis 39: 206-217; Siner, J. et al (2008) "Increased serum angiopoietin 2 levels are associated with increased mortality in sepsis" Shock Sep 11, Epub ahead of print; Sowunmi, A. (1993), "Misdiagnosis of cerebral malaria in adolescents and adults in an endemic area". Trans Royal Soc Trop Med Hyg. 88, 493; Suprin et al. (2000), "Procalcitonin: a valuable indicator of infection in a medical ICU?" Int Care Med 26: 1232-1238; Thwing, J. et al. (2007), "Malaria surveillance - United States, 2005". MMWR Surveill Summ 56(6): 23-40; Ugarte, H. et al. (1999), "Procalcitonin used as a marker of infection in the intensive care unit". Crit Care Med 27: 498-504; Uzzan, B. et al. (2006), "Serum PCT in uncomplicated falciparum malaria. A preliminary study". Trav Med & InfDis March 2006, pp 77-80; Uzzan, B. et al. (1998), Crit Care Med 34(7): 1996-2003; Vicas, A. (2005), "Imported malaria at an inner-city hospital in the United States". Am L Med Sci 329(1): 6-12; and Yeo, T. et al. (2008) "Angiopoietin 2 is associated with decreased endothelial nitric oxide and poor clinical outcome in severe falciparum malaria". PNAS 105(44): 17097-17102.

[0270] This concludes the description of presently preferred embodiments of the invention. The foregoing description has been presented for the purpose of illustration and is not intended to be exhaustive or to limit the invention to the precise form disclosed. Other modifications, variations and alterations are possible in light of the above teaching and will be apparent to those skilled in the art, and may be used in the design and manufacture of other embodiments according to the present invention without departing from the spirit and scope of the invention. It is intended the scope of the invention be limited not by this description but only by the claims forming a part of this application and/or any patent issuing herefrom.

Claims

WHAT IS CLAIMED IS:

1. A rapid diagnostic test panel for use in the diagnosis of malaria and severe bacterial infection in a test sample, wherein the panel comprises monoclonal antibodies to: (a) histidine- rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P .falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2; and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin.

2. A panel according to claim 1, wherein P. falciparum aldolase is the first group member, Angiopoietin-2 is the second group member, and procalcitonin is the third group member.

3. A kit for use in the diagnosis of malaria and severe bacterial infection in a test sample, wherein the kit comprises, in package form, monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P. falciparum aldolase and pan P. falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin.

4. A kit according to claim 3, wherein P. falciparum aldolase is the first group member, Angiopoietin-2 is the second group member, and procalcitonin is the third group member.

5. A rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample, wherein the device comprises: a receiving portion adapted to receive a panel which includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P. falciparum aldolase and pan P.falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin; a sample chamber adapted to receive the sample for contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member; a detection element for detecting signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member; and one or more processors operatively encoded with an algorithm for analysis of the signals to, on the basis of the reactions detected, automatically generate a test result which differentiates between (i) P.falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P.falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria, (iii) non-Z*. falciparum malaria, (iv) infection which is likely to be said severe bacterial infection, and (v) infection which is unlikely to be said severe bacterial infection, in the sample.

6. A device according to claim 5, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and wherein when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or above a predetermined second group high cut-off level, the processors generate the test result as indicative of said P.falciparum malaria which is likely to be at risk for said severe or cerebral malaria.

7. A device according to claim 5, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and wherein when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or below a predetermined second group low cut-off level, the processors generate the test result as indicative of said P .falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria.

8. A device according to any one of claims 5 to 7, wherein when the reactions are detected with the monoclonal antibodies to the first group member but not with the monoclonal antibodies to HRP2, the processors generate the test result as indicative of said non-P. falciparum malaria.

9. A device according to any one of claims 5 to 8, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and wherein when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or above a predetermined third group high cut-off level, the processors generate the test result as indicative of said infection which is likely to be said severe bacterial infection.

10. A device according to any one of claims 5 to 8, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and wherein when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or below a predetermined third group low cut-off level, the processors generate the test result as indicative of said infection which is unlikely to be said severe bacterial infection.

11. A device according to any one of claims 5 to 10, wherein the algorithm operatively encodes the processors to, when the processors generate the test result as indicative of said infection which is unlikely to be said severe bacterial infection, also generate the test result as indicative of a localized bacterial infection and/or a viral infection.

12. A device according to any one of claims 5 to 11, further comprising a presentation element adapted for presentation to a user of treatment and/or follow-up information related to the test result.

13. A device according to claim 5, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and further comprising a presentation element adapted for presentation to a user of treatment and/or follow-up information when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified between a predetermined second group high cut-off level and a predetermined second group low cut-off level.

14. A device according to any one of claims 5 to 8, wherein the algorithm operatively encodes the processors to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and further comprising a presentation element adapted for presentation to a user of treatment and/or follow-up information when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified between a predetermined third group high cut-off level and a predetermined third group low cutoff level.

15. A device according to any one of claims 5 to 14, adapted for use with P. falciparum aldolase as the first group member, Angiopoietin-2 as the second group member, and procalcitonin as the third group member.

16. A device according to one of claims 6 and 13, wherein the second group high cut-off level is predetermined based on local normative data.

17. A device according to one of claims 7 and 13, wherein the second group low cut-off level is predetermined based on local normative data.

18. A device according to one of claims 9 and 14, wherein the third group high cut-off level is predetermined based on local normative data.

19. A device according to any one of claims 9, 14 and 18, adapted for use with procalcitonin as the third group member, and wherein the third group high cut-off level is about 2.0 nanograms per millilitre (ng/mL).

20. A device according to one of claims 10 and 14, wherein the third group low cut-off level is predetermined based on local normative data.

21. A device according to any one of claims 10, 14 and 20, adapted for use with procalcitonin as the third group member, and wherein the third group low cut-off level is about 0.5 nanograms per millilitre (ng/mL).

22. A device according to any one of claims 5 to 21, wherein the sample chamber is adapted either to contact the sample with the monoclonal antibodies inside the sample chamber, or to flow the sample into contact with the monoclonal antibodies outside of the sample chamber.

23. A device according to any one of claims 5 to 22, wherein the device is a handheld portable rapid diagnostic test device adapted for use at a point-of-care.

24. A device according to any one of claims 5 to 23, wherein the signals are light signals, and wherein the detection element is an optical detection element.

25. A device according to any one of claims 5 to 24, further comprising an onboard receiver element adapted to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database, and wherein the algorithm automatically generates the test result at least in part on the basis of the clinical, epidemiological and/or sociological data.

26. A device according to any one of claims 5 to 25, further comprising an onboard transmission element adapted to remotely transmit the test result for storage in a remote database.

27. A device according to any one of claims 5 to 25, further comprising an onboard geolocalization element adapted to generate a geographic location result based on the geographic location of the device; and an onboard transmission element adapted to remotely transmit the test result together with the geographic location result for storage in a remote database.

28. A device according to any one of claims 5 to 27 adapted for use with whole blood, plasma and/or serum as the sample.

29. A method of operating a rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample, wherein the method comprises the steps of: loading a receiving portion of the test device with a panel which includes monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P .falciparum aldolase and pan P .falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin; loading the sample into a sample chamber of the test device; using the test device to contact the sample with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member; using a detection element of the test device to detect signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member; using one or more processors onboard the rapid diagnostic test device to analyze the signals and automatically differentiate between (i) P. falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P .falciparum malaria which is unlikely to be at risk for severe or cerebral malaria, (iii) non-P. falciparum malaria, (iv) infection which is likely to be a severe bacterial infection, and (v) infection which is unlikely to be a severe bacterial infection, in the sample.

30. A method according to claim 29, wherein the processors are used to quantify or semi- quantify the reactions with the monoclonal antibodies to the second group member; and wherein when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or above a predetermined second group high cut-off level, the processors are used to generate the test result as indicative of said P .falciparum malaria which is likely to be at risk for said severe or cerebral malaria.

31. A method according to claim 29, wherein the processors are used to quantify or semi- quantify the reactions with the monoclonal antibodies to the second group member; and wherein when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or below a predetermined second group low cut-off level, the processors are used to generate the test result as indicative of said P. falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria.

32. A method according to any one of claims 29 to 31, wherein when the reactions are detected with the monoclonal antibodies to the first group member but not with the monoclonal antibodies to HRP2, the processors are used to generate the test result as indicative of said non- P. falciparum malaria.

33. A method according to any one of claims 29 to 32, wherein the processors are used to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and wherein when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or above a predetermined third group high cut-off level, the processors are used to generate the test result as indicative of said infection which is likely to be said severe bacterial infection.

34. A method according to any one of claims 29 to 32, wherein the processors are used to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and wherein when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or below a predetermined third group low cut-off level, the processors are used to generate the test result as indicative of said infection which is unlikely to be said severe bacterial infection.

35. A method according to any one of claims 29 to 34, further comprising a step of using the processors to, when said infection is differentiated as unlikely to be severe bacterial infection, automatically characterize the sample as indicating localized bacterial and/or a viral infection.

36. A method according to any one of claims 29 to 35, further comprising a step of using a presentation element onboard the test device to present treatment and/or follow-up information related to the test result to a user.

37. A method according to claim 29, wherein the processors are used to quantify or semi- quantify the reactions with the monoclonal antibodies to the second group member; and further comprising a step of using a presentation element onboard the test device to present treatment and/or follow-up information to a user when the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified between a predetermined second group high cut-off level and a predetermined second group low cut-off level.

38. A method according to any one of claims 29 to 32, wherein the processors are used to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and further comprising a step of using a presentation element onboard the test device to present treatment and/or follow-up information to a user when the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified between a predetermined third group high cut-off level and a predetermined third group low cut-off level.

39. A method according to any one of claims 29 to 38, adapted for use with P .falciparum aldolase as the first group member, Angiopoietin-2 as the second group member, and procalcitonin as the third group member.

40. A method according to one of claims 30 and 37, further comprising a step of predetermining the second group high cut-off level based on local normative data.

41. A method according to one of claims 31 and 37, further comprising a step of predetermining the second group low cut-off level based on local normative data.

42. A method according to one of claims 33 and 38, further comprising a step of predetermining the third group high cut-off level based on local normative data.

43. A method according to any one of claims 33, 38 and 42, adapted for use with procalcitonin as the third group member, and wherein the third group high cut-off level is about 2.0 nanograms per millilitre (ng/mL).

44. A method according to one of claims 34 and 38, further comprising a step of predetermining the third group low cut-off level based on local normative data.

45. A method according to any one of claims 34, 38 and 44, adapted for use with procalcitonin as the third group member, and wherein the third group low cut-off level is about 0.5 nanograms per millilitre (ng/mL).

46. A method according to any one of claims 29 to 45, the test device is used to contact the sample with the monoclonal antibodies either inside or outside of the sample chamber.

47. A method according to any one of claims 29 to 46, adapted for use at a point-of-care with a handheld portable device as the test device.

48. A method according to any one of claims 29 to 47, wherein the detection element is used to detect light signals as the signals.

49. A method according to any one of claims 29 to 48, further comprising a step of using a receiver element onboard the test device to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database, and wherein the processors are used to generate the test result at least in part on the basis of the clinical, epidemiological and/or sociological data.

50. A method according to any one of claims 29 to 49, further comprising a step of using a transmission element onboard the test device to remotely transmit the test result for storage in a remote database.

51. A method according to any one of claims 29 to 49, further comprising a step of using a geolocalization element onboard the test device to generate a geographic location result based on the geographic location of the device; and a step of using a transmission element onboard the test device to remotely transmit the test result together with the geographic location result for storage in a remote database.

52. A method according to any one of claims 29 to 51 adapted for use with whole blood, plasma and/or serum as the sample.

53. A computer readable medium on which is stored instructions which upon execution will operate a rapid diagnostic test device for use in the diagnosis of malaria and severe bacterial infection in a test sample, wherein the instructions comprise: instructions for contacting a panel received by the test device with the sample received within a sample chamber of the test device, with the panel including monoclonal antibodies to: (a) histidine-rich protein II (HRP2), (b) at least one first group member selected from a first group consisting of P .falciparum aldolase and pan P '.falciparum lactate dehydrogenase, (c) at least one second group member selected from a second group consisting of Angiopoietin-1 and Angiopoietin-2, and (d) at least one third group member selected from a third group consisting of C-reactive protein and procalcitonin; instructions for using a detection element of the test device to detect signals emitted from, or absorbed by, the sample in contact with the monoclonal antibodies to HRP2, to the first group member, to the second group member and to the third group member; and instructions for one or more processors onboard the test device to analyze the signals and automatically differentiate between (i) P. falciparum malaria which is likely to be at risk for severe or cerebral malaria, (ii) P .falciparum malaria which is unlikely to be at risk for severe or cerebral malaria, (iii) non- P.falciparum malaria, (iv) infection which is likely to be a severe bacterial infection, and (v) infection which is unlikely to be a severe bacterial infection, in the sample.

54. A computer readable medium according to claim 53, wherein the instructions for the processors include instructions: to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and to generate the test result as indicative of said P. falciparum malaria which is likely to be at risk for said severe or cerebral malaria when (a) the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and (b) the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or above a predetermined second group high cut-off level.

55. A computer readable medium according to claim 53, wherein the instructions for the processors include instructions: to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and to generate the test result as indicative of said P. falciparum malaria which is unlikely to be at risk for said severe or cerebral malaria when (a) the reactions are detected with the monoclonal antibodies to HRP2 and to the first group member, and (b) the reactions with the monoclonal antibodies to the second group member are quantified or semi-quantified at or below a predetermined second group low cut-off level.

56. A computer readable medium according to any one of claims 53 to 55, wherein the instructions for the processors include instructions to generate the test result as indicative of said non-P. falciparum malaria when the reactions are detected with the monoclonal antibodies to the first group member but not with the monoclonal antibodies to HRP2.

57. A computer readable medium according to any one of claims 53 to 56, wherein the instructions for the processors include instructions: to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and to generate the test result as indicative of said infection which is likely to be said severe bacterial infection when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and (b) the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or above a predetermined third group high cut-off level.

58. A computer readable medium according to any one of claims 53 to 56, wherein the instructions for the processors include instructions: to quantify or semi-quantify the reactions with the monoclonal antibodies to the third group member; and to generate the test result as indicative of said infection which is unlikely to be said severe bacterial infection when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and (b) the reactions with the monoclonal antibodies to the third group member are quantified or semi-quantified at or below a predetermined third group low cut-off level.

59. A computer readable medium according to any one of claims 53 to 58, wherein the instructions for the processors include instructions to automatically characterize the sample as indicating localized bacterial and/or a viral infection when said infection is differentiated as unlikely to be severe bacterial infection.

60. A computer readable medium according to any one of claims 53 to 59, wherein the stored instructions also include instructions for using a presentation element onboard the test device to present treatment and/or follow-up information related to the test result to a user.

61. A computer readable medium according to claim 53, wherein the instructions for the processors include instructions to quantify or semi-quantify the reactions with the monoclonal antibodies to the second group member; and wherein the stored instructions also include instructions for using a presentation element onboard the test device to present treatment and/or follow-up information to a user when (a) the reactions are detected with the monoclonal antibodies io HRP2 and to the first group member, and (b) the reactions with the monoclonal antibodies to the second group member a/e quantified or semi-quamified between a predetermined second group high cut-off level and a predetermined second group low cut-off fevel.

62. A computer readable medium according Lo un> one of claims 53 to 56, wherein the instructions for the processors include instructions to quantify or scmi-quantify the reactions with the monoclonal antibodies to the third group member; ;md wherein the stored instructions also include instructions for using a presentation element onboard the test device to present treatment and/or follow-up information to a user when (a) the reactions are detected with neither the monoclonal antibodies to HRP2 nor to the first group member, and (b) the reactions with the monoclonal antibodies to the third group member arc quantified or semi-quantified between a predetermined third group high cut-off level and a prcdeiermined third group low cut-off level.

63. A computer readable medium accoκliπg to any one of claims 53 to 62, wherein the stored instructions arc adapted for use with P falciparum aldolase as the first βroup member, Angioρoietin-2 as the second group member, and procaloitonin as the third group member.

64. A computer readable medium accoiding to one of claims 57 and 62, wherein the stored instructions are adapted for use with procalciumin as the third βroup member, and wherein the third group high cut-off level is about 2.0 nanograms per millilitre (nβ/mL),

65. A computer readable medium according Io one of claims 58 and 62, wherein the stored instructions are adapted for use wiih procalcitonin as the third group member, and wherein the third group low cut-off level is about 0.5 nanograms pω millilitre (ng/mL).

66. A computer readable medium according to any one of claims 53 to 65, wherein the stored instructions also include instructions for using a receiver element onboard the test device to remotely receive clinical, epidemiological and/or sociological data regarding the sample from a remote database; and wherein the instructions for the processors include instructions to generate the test result at least in part on the basis of the clinical, epidemiological and/or sociological data.

67. A computer readable medium according to any one of claims 53 to 66, wherein the stored instructions also include instructions for using a transmission element onboard the test device to remotely transmit the test result for storage in a remote database.

68. A computer readable medium according to any one of claims 53 to 66, wherein the stored instructions also include instructions for using a geolocalization element onboard the test device to generate a geographic location result based on the geographic location of the device; and for using a transmission element onboard the test device to remotely transmit the test result together with the geographic location result for storage in a remote database.