WO2003050556A2 - Dispositif et procede d'analyse - Google Patents

Dispositif et procede d'analyse Download PDF

Info

Publication number
WO2003050556A2
WO2003050556A2 PCT/GB2002/005644 GB0205644W WO03050556A2 WO 2003050556 A2 WO2003050556 A2 WO 2003050556A2 GB 0205644 W GB0205644 W GB 0205644W WO 03050556 A2 WO03050556 A2 WO 03050556A2
Authority
WO
WIPO (PCT)
Prior art keywords
binding partner
specific binding
labelled
target moiety
detection zone
Prior art date
Application number
PCT/GB2002/005644
Other languages
English (en)
Other versions
WO2003050556A3 (fr
Inventor
Michael Frederick Sanders
Original Assignee
The Secretary Of State For Environment, Food & Rural Affairs
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Secretary Of State For Environment, Food & Rural Affairs filed Critical The Secretary Of State For Environment, Food & Rural Affairs
Priority to AU2002350951A priority Critical patent/AU2002350951A1/en
Publication of WO2003050556A2 publication Critical patent/WO2003050556A2/fr
Publication of WO2003050556A3 publication Critical patent/WO2003050556A3/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54306—Solid-phase reaction mechanisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/32—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Bacillus (G)

Definitions

  • the present application relates to a method for conducting an assay for a target moiety such as a microorganism, which is present at relatively high concentrations in a sample, as well as to assay devices and kits for use in the method.
  • Such techniques are only applicable however where the moiety being detected can be mobilised relatively easily, so that it can flow through pores . in the membrane . Thus such methods are less useful where the moiety to be detected is large, such as microorganisms or microorganism spores. Furthermore, high levels of bacteria in a sample may also inhibit such tests, making dilutions of sample necessary before the test can be operated successfully. Although generally bacteria will be present in most samples at relatively low concentrations, there are instances, in particular where plant pathogens and particularly food spoilage organisms are concerned, where they may present in very high concentrations . Examples of such organisms include bacteria from the Erwinia, Pseudomonas and Xanthomonas strains .
  • microorganisms and the like may be carried out in homogenous assays such as those described in US Patent No. 5888725.
  • a problem commonly encountered in such assays is that the concentration of the organism in a sample is generally very low. Thus it may be necessary to incubate the sample for a significant period of time prior to conducting the assay in order to culture the organism to detectable levels. This delay may be unacceptable in many situations where public health is at risk, for example, where biological warfare or terrorism is suspected.
  • PCR polymerase chain reaction
  • some organisms such as those that form spores (e.g. Bacillus anthracls) , may be difficult to break open to liberate detectable cellular components. In such cases, it may be necessary to grow out the spore to provide vegetative cells for PCR or other molecular biological methods, which will increase the time taken to obtain the results, potentially to an unacceptable level.
  • Bacillus anthracls The rapid detection Bacillus anthracls is currently very important in the field of public safety and likely to remain so for some considerable period of time. Bacillus anthracls has been investigated for its potential as a biological warfare agent and possibly used as such for some time. More recently it has been used as a terrorist weapon and has been targeted towards the largely unprotected civilian population.
  • a method for detecting the presence of a target moiety in a sample comprising: a) immersing in a liquid sample suspected of containing said moiety, a detection zone on a solid support, said detection zone having a first specific binding partner, which specifically binds said target moiety, immobilised therein, said first specific binding partner being present in the detection zone an amount which binds up to a predetermined concentration of said target moiety, b) removing said solid support from the sample, c) subsequently immersing the detection zone on the solid support in a solution or suspension of a labelled second specific binding partner which binds said first specific binding partner, wherein the amount of said labelled second specific binding partner is at least sufficient to bind to all said first binding partner, d) detecting the presence of labelled material in the detection zone on said solid support; and e) relating the result of (d) to the presence of target moiety in said sample.
  • the method of the invention is simple to operate and should give rapid results in situations
  • any of the target moiety present binds to the first specific binding partner in the detection zone, blocking at least some of the first specific binding partner from further binding. Where there is sufficient target moiety, all of the binding sites in the detection zone on the support will be occupied in this way.
  • the solid support with the target moiety bound thereto is then separated from the sample and subsequently immersed in a second liquid containing a labelled second specific binding partner, which binds the first specific binding partner.
  • the assay may therefore be operated in a semi-quantitative fashion by detecting the reduction in the signal and thus the reduction in the amount of labelled second specific binding partner which may be bound to the solid support, as a result of exposure to the sample. This is best achieved by calibrating the assay device with known quantities of labelled second binding partner.
  • the method of the invention can be suitably applied to the detection of a moiety for which a specific binding partner may be identified.
  • a specific binding partner may be identified.
  • it may be used to detect proteins, polypeptides or microorganisms including bacteria, bacterial spores and viruses (for which specific binding partners may be antibodies or binding fragments, or receptors or the like) as well as nucleic acid sequences such as DNA and RNA (where the first specific binding partner will comprise complementary nucleic acid sequences which hybridise to the target sequence) .
  • the method is suitably applied to the detection of relatively large moieties, such as microorganisms, and in particular bacteria and bacterial spores .
  • they may comprise anthrax spores such as those used in terrorist attacks, where powders containing the spores have been delivered through the mail .
  • the may comprise plant pathogens, which may be present in relatively high concentrations in infected plant tissue,
  • pathogens may be viruses, but in particular are bacteria such as Erwlnla , Pseudomonas and Xanthomonas species.
  • a troseptlca which causes Black Heart and Soft Rot in particular in potato (Solanum tuberosum) ; Erwlnla carotovora subsp. carotovora which causes bacterial Soft rot in particular in shallot ⁇ Alllum ascalonlcum) , onion (Alllum cepa) , Alllum sp., broccoli, cauliflower (Brasslca oleracea var. botrytls) and cabbage ⁇ Brasslca oleracea var.
  • Erwlnla chrysantheml which causes Bacterial Blight in florist's chrysanthemum ( Chrysanthemum morlfollum) ; Erwlnla herblcola which causes Leafspot in syngonlum ( Syngonlum podophyllum) , Pink Disease in pineapple (Ananas comosus) and Purple Stain; Erwlnla Sp. which affects watermelon (Cltrullus vulgaris) and also Fire blight of apple, although this may be caused by Erwlnla amylovora .
  • bacteria include Streptococcus pneumonias, Nelsserla menlngltldls, Salmonella sp, Staphylococcus aureus, Haemophllus Influenzae type b(Hib) Escherlchla coll (particularly those strains containing the Kl polysaccharide) , Llsterla monocytogenes Enterococcl, nonenterococcal group D streptococci, -he olytic streptococci, and other gram-negative enteric organisms (eg, Klebslella sp, Enterobacter sp. and Cltrobacter dlversus)
  • Streptococcus pneumonias Nelsserla menlngltldls
  • Salmonella sp Staphylococcus aureus
  • Haemophllus Influenzae type b(Hib) Escherlchla coll particularly those strains containing the Kl polysaccharide
  • Bacteremia and septic shock are closely related conditions .
  • Bacteremia denotes bacteria in the bloodstream.
  • Septic shock is sepsis with hypoperfusion and hypotension refractory to fluid therapy.
  • Sepsis refers to a serious infection, localized or bacteremic, that is accompanied by systemic manifestations of inflammation. Sepsis due to bacteremia is often called septicemia; this often imprecisely used term is now being discouraged.
  • the more general term, systemic inflammatory response syndrome recognises that several severe conditions (e.g., infections, pancreatitis, burns, trauma) can trigger an acute inflammatory reaction, the systemic manifestations of which are associated with release into the bloodstream of a large number of endogenous mediators of inflammation. Diagnosis depends on isolating bacteria from blood and avoiding contamination with bacteria from the skin.
  • Sample may suitably be prepared by taking a sample of any suspect material such as powder, or contaminated plant or other organism, and suspending this in water or an aqueous liquid.
  • This liquid may contain suspending agents, surfactants or detergents if necessary to achieve good suspension of the target moiety.
  • this may be centrifuged or filtered if necessary to provide a usable serum sample.
  • the concentration of first binding agent on the " support it would be possible to set the concentration of first binding agent on the " support to be such that any concentration above levels which are known to- cause infection (i.e. more than 10 4 spores per sample) will give a 100% positive result (zero signal) .
  • the test will provide a rapid method for detecting risk to persons exposed to any suspect powder or the like, and will allow antibiotic treatment to begin immediately.
  • the first specific binding partner suitably comprises an antibody or a binding fragment thereof, which is specific for the target moiety such as- the microorganism.
  • Antibodies may be monoclonal or polyclonal .
  • the solid support is suitably in the form of a single self- supporting unit such as a slide or membrane, for instance, a nitrocellulose membrane, which can be readily moved from one sample tube to another and if necessary also into a detector, either manually or using an automated device. It is suitably prepared by immobilising suitable first specific binding partners thereon using conventional methods, such as by spraying or covalent or other bonding, in a detection zone. The amount of first specific binding partner applied in this way is suitably measured to ensure that similar amounts are applied to similar batches of the assay device.
  • any remaining binding sites on the support are suitably blocked by immersion in a blocking solution as is known in the art.
  • a blocking solution as is known in the art.
  • unused sites on nitrocellulose membranes may be blocked with polyvinylalcohol as described in GB-A-2204398.
  • the device is suitably a dipstick type device comprising a nitrocellulose membrane, if necessary on a backing support to make it self-supporting, or within a protective housing or cover such as are well known in the art.
  • the construction of the support must be such however that the detection zone containing the immobilised first binding agent is immersible in the sample solution or suspension. This area should then be accessible for detecting signal, for example by being visible to the eye, or for inclusion in a detection device such as a fluorimeter or luminometer .
  • the support is subject to washing between steps (b) and (C) and/or (c) and (d) .
  • Step (c) of the method of the invention is suitably effected by immersing the detection zone on the solid support into a solution or suspension of the second specific binding partner so as to allow any available first specific binding partner to bind to it.
  • the nature of the labelled second specific binding partner will vary depending upon the nature of the target moiety and the first specific binding partner.
  • the labelled second specific binding partner will comprise a labelled cell or spore. Where these comprise a pathogenic organism, the cell or spore is suitably attenuated or inactivated.
  • the labelled second specific binding partner will comprise a labelled protein in particular an antigen which binds the first specific binding partner.
  • such forms will comprise a microsphere coated with the antigen.
  • the microsphere is conveniently a labelled microsphere of a size similar to that of the microorganism or spore which forms the target moiety, so that it will bind in a similar fashion to that of the microorganism or spore and thus mimic the binding of the target moiety.
  • a wide variety of microspheres for example hydrophobic polymeric microspheres are available for example from Bangs Labs . These include florescently labelled microspheres .
  • the label used in the labelled second specific binding partner is a visible label, such as a fluorescent label, or a chemiluminescent label as well as coloured particulate labels such as coloured latex particles, or gold or silver particles.
  • labels such as those which need to be “developed” may be employed. These will include enzymes used in conventional ELISA tests, such as horse-radish peroxidase and phosphatase, which form coloured soluble reaction products on contact with the enzyme substrate.
  • enzymes used in conventional ELISA tests such as horse-radish peroxidase and phosphatase, which form coloured soluble reaction products on contact with the enzyme substrate.
  • the label is a fluorescent or chemiluminescent label, which may be detected using a fluorimeter or luminometer, capable of detecting the intensity of the signal generated.
  • Fluorescent labels are available from a variety of sources and include fluorescein, SybrgreenTM, SybrGold TM, Cy5, rhodamine dyes, as well as pyridyloxazole dyes such as Cascade yellow and dapoxyl dyes and dyes based on the 6,8- difluoro-7-hydroxycoumarin fluorophore such as Marina blue and Pacific blue dyes available from Molecular Probes .
  • the results obtained in step (d) of the method of the invention will be compared with results obtained using known concentrations of labelled second binding partner.
  • Such results are suitably formulated into a calibration graph. In this way any non-specific binding of the labelled second binding partner to the support would be detected and this could act as the baseline for the results.
  • detectable levels between the baseline and 100% saturation of the indicator would provide an indication of the quantity of target moiety present.
  • the method is based on a dipstick type of assay, which is portable and easy to use.
  • a substrate such as a dipstick or slide would be covered with stable antibody to for example Bacillus anthracls spores.
  • this substrate would be so engineered as to contain a fixed number of binding sites and as such only able to bind a finite number of spores.
  • a preparation of biologically inactive Bacillus anthracls spores would be available. These inactive spores would be labelled with a suitable marker (luminescent or fluorescent for example) .
  • kits provide a further aspect of the invention.
  • the invention further provides a kit for conducting a method as described above, said kit comprising a solid support having immobilised thereon in a detection zone, a predetermined amount of a first specific binding partner which binds a target moiety, said detection zone being arranged such that it is immersible in a liquid sample, and an excess of a labelled second specific binding partner which binds said first binding partner and which mimics said target moiety.
  • suspending agents such as suspending agents, detergents or surfactants, as well as suitable containers for' sample and/or suspension of second specific binding partner may also be included in the kit.
  • the method and kits of the invention can be used as a preliminary assessor of risk, to allow early treatment to begin, where positive results are found.
  • the test results obtained in this way may then be confirmed or otherwise using for example laboratory based tests, or PCR methods, which may take too long in some instances .
  • Figure 1 illustrates schematically, the assay of the invention wherein the sample contains a high concentration of analyte
  • Figure 2 illustrates schematically, the assay of the invention wherein the sample contains a moderate concentration of analyte.
  • An antibody able to bind a target bacterial cell or spore is applied to a substrate in the form of a slide or dipstick, able to be inserted into a measuring device.
  • the measuring device is suitably an instrument containing a UV or visible light source designed to deliver a wavelength able to excite a dye or other marker.
  • the instrument will also contain a detector able to measure the light emitted from the excited dye .
  • the substrate is then calibrated with either inactivated bacterial spores or cells labelled with fluorescent dye or fluorescently labelled microspheres coated with the target antigen.
  • Substrates are prepared and calibrated in batches and will be designed to be consistent within each batch. Each substrate will be labelled to saturation point, washed and measured. This result will give the 100% saturation/detection value- of the signal.
  • blank antibody-labelled substrates are exposed to the target organism or spore in suspension.
  • the substrate is washed and exposed to same, labelled substance described above in relation to the calibration process.
  • the substrate After washing the substrate is placed in a detector and the signal at the excitation wavelength of the label or dye is measured. Any signal less that the 100% saturation signal level or other predetermined level will indicate the presence of the target due to blocking of the binding sites .
  • FIG. 1 An assay of this type is illustrated schematically in Figures 1 and 2.
  • a solid substrate (1) has immobilised thereon in a detection zone, a quantity of antibody (2) that is specific for a target analyte such as a bacterial sport.
  • the substrate is calibrated as described above, so that the concentration of analyte, which will bind to it, is known.
  • the substrate (1) is then immersed in a sample (3) which includes a relatively high concentration of bacterial spores (4) in suspension in water.
  • the substrate is maintained in the sample for a sufficient period of time to allow the spores to bind to the antibodies (2) .
  • all the available sites within the detection zone on the substrate (1) become blocked.
  • the substrate is removed from the sample 3, if necessary washed, and subsequently immersed in a solution of labelled inactivated spores (6). These spores would also bind the antibodies (2), if there were any available for binding. However, in this instance, all the binding sites have been blocked. When the substrate is removed from this solution, no signal from the label (6) is visible. It may be necessary to insert the substrate into a signal reading device such as a fluorimeter or luminometer as described above.
  • the concentration of the spores in the sample (3) is above the level set by the calibration of the substrate. In particular, it may be above a level where infection results, and therefore, prophylactic or therapeutic treatments may be administered to anyone who has been exposed to the contents of the sample.'
  • the sample contains less spores (4) , which means that after incubation and removal of the substrate from the sample, some antibody sites (2) on the substrate (1) are still available. As a result, immersion in the second solution results in the adherence of some labelled material (6) to the available sites. These will be detected in the signal reading device.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de détection de la présence d'un groupe fonctionnel cible dans un échantillon, consistant a) à immerger une zone de détection située sur un support solide, dans un échantillon liquide susceptible de contenir ledit groupe fonctionnel, ladite zone de détection comportant un premier partenaire de liaison spécifique liant spécifiquement ledit groupe fonctionnel cible, immobilisé dans celle-ci, et ledit partenaire de liaison spécifique étant présent dans la zone de détection à une quantité permettant de lier ledit groupe fonctionnel cible à une concentration prédéterminée ; b) à extraire ledit support solide de l'échantillon ; c) à immerger ensuite la zone de détection située sur le support solide dans une solution ou une suspension d'un deuxième partenaire de liaison spécifique marqué liant ledit premier partenaire de liaison spécifique, la quantité dudit deuxième partenaire de liaison spécifique marqué suffisant à lier l'ensemble dudit premier partenaire de liaison ; d) à détecter la présence de l'élément marqué dans la zone de détection située sur ledit support solide ; et, e) à déterminer la présence du groupe fonctionnel cible dans ledit échantillon en fonction du résultat de l'étape (d). Le procédé selon l'invention sert notamment de test de détection rapide d'organismes pathogènes tels que des spores d'anthrax pouvant être présents dans des concentrations élevées.
PCT/GB2002/005644 2001-12-13 2002-12-13 Dispositif et procede d'analyse WO2003050556A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002350951A AU2002350951A1 (en) 2001-12-13 2002-12-13 Assay device and method

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0129776A GB0129776D0 (en) 2001-12-13 2001-12-13 Assay device and method
GB0129776.1 2001-12-13

Publications (2)

Publication Number Publication Date
WO2003050556A2 true WO2003050556A2 (fr) 2003-06-19
WO2003050556A3 WO2003050556A3 (fr) 2003-10-16

Family

ID=9927520

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2002/005644 WO2003050556A2 (fr) 2001-12-13 2002-12-13 Dispositif et procede d'analyse

Country Status (3)

Country Link
AU (1) AU2002350951A1 (fr)
GB (2) GB0129776D0 (fr)
WO (1) WO2003050556A2 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005026393A2 (fr) * 2003-02-17 2005-03-24 Midwest Research Institute, Inc. Analogue de matiere particulaire biologique
WO2013016449A2 (fr) * 2011-07-26 2013-01-31 Indicator Systems International, Inc. Essais pour la détection de microbes
CN111735954A (zh) * 2020-06-23 2020-10-02 南京农业大学 梨火疫病菌快速免疫检测试纸条及其应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2796859A1 (fr) * 2013-04-25 2014-10-29 Biocartis SA Quantification d'une surface fonctionnalisée

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4279885A (en) * 1978-05-19 1981-07-21 Becton Dickinson & Company Solid phase assay
US4391795A (en) * 1980-03-21 1983-07-05 Becton Dickinson And Company Assay for free thyroid hormone
EP0267317A1 (fr) * 1983-05-20 1988-05-18 Profile Diagnostic Sciences Inc. Procédé pour la détection des protéines et virus
WO1988008978A1 (fr) * 1987-05-14 1988-11-17 The Mclean Hospital Corporation Immuno-analyse d'antigenes multiples
EP0311492A2 (fr) * 1987-09-30 1989-04-12 Elf Sanofi Trousse et méthode de dosage immunométrique applicables à des cellules entières
WO1990015327A1 (fr) * 1989-06-06 1990-12-13 Martin Gould Kit diagnostic d'immunoanalyse
EP0577092A2 (fr) * 1992-07-02 1994-01-05 Becton, Dickinson and Company Immunoessai utilisant des particules contenant des substances détectables différentes
WO2001049823A2 (fr) * 2000-01-06 2001-07-12 Biosite Diagnostics, Inc. Dosage pour la detection de bacillus anthracis
WO2001083561A2 (fr) * 2000-04-28 2001-11-08 Tetracore, L.L.C. Anticorps spécifiques de l'anthrax

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU6479986A (en) * 1985-10-22 1987-05-19 Murex Corp. Idiotypic-antigenic conjunction binding assay
US6316205B1 (en) * 2000-01-28 2001-11-13 Genelabs Diagnostics Pte Ltd. Assay devices and methods of analyte detection

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4279885A (en) * 1978-05-19 1981-07-21 Becton Dickinson & Company Solid phase assay
US4391795A (en) * 1980-03-21 1983-07-05 Becton Dickinson And Company Assay for free thyroid hormone
EP0267317A1 (fr) * 1983-05-20 1988-05-18 Profile Diagnostic Sciences Inc. Procédé pour la détection des protéines et virus
WO1988008978A1 (fr) * 1987-05-14 1988-11-17 The Mclean Hospital Corporation Immuno-analyse d'antigenes multiples
EP0311492A2 (fr) * 1987-09-30 1989-04-12 Elf Sanofi Trousse et méthode de dosage immunométrique applicables à des cellules entières
WO1990015327A1 (fr) * 1989-06-06 1990-12-13 Martin Gould Kit diagnostic d'immunoanalyse
EP0577092A2 (fr) * 1992-07-02 1994-01-05 Becton, Dickinson and Company Immunoessai utilisant des particules contenant des substances détectables différentes
WO2001049823A2 (fr) * 2000-01-06 2001-07-12 Biosite Diagnostics, Inc. Dosage pour la detection de bacillus anthracis
WO2001083561A2 (fr) * 2000-04-28 2001-11-08 Tetracore, L.L.C. Anticorps spécifiques de l'anthrax

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PHILIPS A P ET AL: "MONOCLONAL ANTIBODIES AGAINST SPORE ANTIGENS OF BACILLUS ANTHRACIS" FEMS MICROBIOLOGY IMMUNOLOGY, ELSEVIER, GB, vol. 47, no. 3, December 1988 (1988-12), pages 169-178, XP001031197 ISSN: 0920-8534 *
PHILLIPS A P ET AL: "EVALUATION OF IMMUNO RADIOMETRIC AND ELISA ENZYME LINKED IMMUNO SORBENT ASSAY VERSIONS OF A MICRO TITER PLATE ASSAY FOR BACILLUS-ANTHRACIS SPORES" JOURNAL OF IMMUNOLOGICAL METHODS, vol. 70, no. 1, 1984, pages 75-82, XP009016145 ISSN: 0022-1759 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005026393A2 (fr) * 2003-02-17 2005-03-24 Midwest Research Institute, Inc. Analogue de matiere particulaire biologique
WO2005026393A3 (fr) * 2003-02-17 2005-06-02 Midwest Res Inst Inc Analogue de matiere particulaire biologique
US7179596B2 (en) 2003-02-17 2007-02-20 Sceptor Industries, Inc. Biological particulate matter analogue
WO2013016449A2 (fr) * 2011-07-26 2013-01-31 Indicator Systems International, Inc. Essais pour la détection de microbes
WO2013016449A3 (fr) * 2011-07-26 2013-04-25 Indicator Systems International, Inc. Essais pour la détection de microbes
US8609355B2 (en) 2011-07-26 2013-12-17 Indicator Systems International, Inc. Assays for the detection of microbes
CN111735954A (zh) * 2020-06-23 2020-10-02 南京农业大学 梨火疫病菌快速免疫检测试纸条及其应用

Also Published As

Publication number Publication date
GB2385123A (en) 2003-08-13
GB0129776D0 (en) 2002-01-30
GB2385123B (en) 2004-05-05
AU2002350951A1 (en) 2003-06-23
WO2003050556A3 (fr) 2003-10-16
GB0229038D0 (en) 2003-01-15

Similar Documents

Publication Publication Date Title
US20040209353A1 (en) Biological sample storage device and method for biological sample contamination testing
EP1687439B1 (fr) Substrats colorimetriques, detecteurs colorimetriques, et leurs procedes d'utilisation
JP2008506930A (ja) 分析物を処理、標識および濃縮するための方法およびデバイス
US11085065B2 (en) Phenotypic engineering of spores
JP4578478B2 (ja) バクテリオファージを使用した微生物を検出するための装置および方法
JP6005053B2 (ja) 微生物の検出および定量方法
EP1613965A2 (fr) Appareil et procede de detection d'organismes vivants microscopiques au moyen de bacteriophage
US20070238145A1 (en) Phenotypic engineering of spores
US8039228B2 (en) Signal amplification using a synthetic zymogen
US6312891B1 (en) Species-specific DNA probes for Vibrio vulnificus and Vibrio cholerae, methods and kits
PL195495B1 (pl) Sposób i zestaw testowy do wykrywania antybiotyków beta-laktamowych w płynach biologicznych
EP1086378B1 (fr) Procede analytique faisant appel a un marquage viral multiple
US20170038373A1 (en) Biosensor using magnetic particles for pathogen detection
Cavalera et al. Development of molecular and antigenic-based rapid tests for the identification of African swine fever virus in different tissues
Zamir et al. Detection of quiescent fungi in harvested fruit using CMOS biosensor: A proof of concept study
WO2003050556A2 (fr) Dispositif et procede d'analyse
Campbell et al. Rapid antigen tests
US20100062418A1 (en) Inactivated and dried biological preparations
US20030059839A1 (en) Method for detecting pathogens using immunoassays
JP2001136999A (ja) 微生物ａｔｐの測定法において添加使用されるａｔｐ消去前処理剤および同処理剤を用いる微生物ａｔｐの高感度測定法
US6524810B2 (en) Method of making bioluminescent assay reagent based on non-viable E. coli
US20240117404A1 (en) Automatic Phagogram
EP3740585B1 (fr) Amélioration de la détection de micro-organismes
JP2002543781A (ja) アッセイ試薬
RU2169925C1 (ru) Способ определения активности лейцинаминопептидазы

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase in:

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP