MXPA99009562A - Equipment, kit and method for microbiological diagnosis - Google Patents

Abstract

The invention concerns microbiology, more specifically equipment, a kit and a method which are used for quick microbiological diagnosis with applications in human and veterinary clinical medicine. The invention enables detection of turbodimetric changes caused by to microbial growth. To this end, equipment using two main devices i.e. a static turbodimetric mini-reader and a microflow sensor actuated by a peristaltic pump which is coupled to a PC with software for acquisition, processing and creation of data bases to produce the necessary reports. The diagnostic kit consists of a flask with a culture medium containing a polymer exhibiting derepression activity and two substrates which can be optionally added for specific identification of E. coli as well as a given amount of antibiotic discs arranged in a strip to determine the antibiogram from isolated colonies or positive samples obtained directly from the sources which contain them, thereby enabling detection of infections of the urinary tract from direct urine samples, including simultaneous identification of E. coli.

Description

EQUIPMENT, GAME AND METHOD FOR THE MICROBIOLOGICAL DIAGNOSIS.

Technical Sector The present invention is related to the branch of microbiology, and in particular to a kit, a set and a method useful in rapid microbiological diagnosis, with application in the human and veterinary clinic. Previous Technique The microbiological diagnosis based on physical, chemical and biological methods is an aspect that has been widely tackled in the previous art. US 8,000,755 The patent relates to an electrochemical method for detecting bacteria by measuring the decrease in polarographic oxygen current flowing through an electroanalytical cell that contains two different electrodes immersed in the inoculated culture medium. This system uses large volumes of culture medium (15-18 ml) for its exploration, which makes it difficult to handle the samples at a routine level. Another method used to detect microbial growth is the principle of the voltaic cell, which is based on the use of an appropriate container with electrodes of noble metals and required volumes, which generate a potential, detecting the loss of said potential in the moment of full growth of the bacteria. In the specialized literature, devices based on this principle have been described. The equipment described in patent GB 83-17685 is of this type and uses a detection method by means of the variation of potential between electrodes that are in contact with the fluid sample, this implicitly involves the measurement of potential low with high impedance plementations. of entrance that causes the affectation of the measurable signal product of undesirable and unavoidable noises in the majority of the cases, besides using container containers with noble metals or non-recoverable gold-plated. An effective and simple method for detecting microbial growth is based on the measurement of the conductimetric changes produced by microbial growth in an appropriate culture medium. According to the pertinent literature, the ionic movements in a solution contained in a conductivity measurement cell produce a signal indicating the conductimetric value of said solution. It is known that the conductivity cells do not have a full linearity in their fullness of scale, and that the measurement depends greatly on the temperature. US Pat. No. 4,482,967 discloses a detector and method for conductivity measurements, which corrects these defects.

This is a high precision and complex equipment, with special requirements to measure absolute values of conductivity in a gas chromatograph, with conventional cells and large volumes. It is known that microbial growth can be detected in fluid samples by different routes, for example, using a turbidimetric method where the growth of bacteria produces a turbidity that is read by the system and compared against known standards. This system requires conventional optical sensors, as well as container containers of the sample of optical quality, complex design to work with samples that include visible solids with the naked eye (eg antibiotic discs). A great disadvantage of this system is that it can not work with interfered samples, that is, it requires optically homogeneous samples, in addition to the optical complexity that it entails. US Patent 3,852,532; US 3,895,661 and US 3,889,011 describe methods and apparatus for these purposes based on these principles. Patents US 4,021,120 and US 3,714,445 describe devices based on the principle of turbidimetry for measuring the growth of microorganisms in liquid media. US Patent 4,021,120 discloses a device for monitoring the growth of microorganisms in a liquid medium containing gas. From the container in which the medium is located, samples are taken with the help of a pump to take them to a degassing chamber in order to eliminate gas bubbles, then the sample is introduced into a measuring chamber where a ray of light is passed through it, which will affect a photoelectric cell, producing a current that is increased by an amplifier, which is an indicator of the growth of the microorganisms. The magnitude of this current will depend on the intensity of the light beam and, in turn, will be influenced by the turbidity of the medium. The sample, once analyzed, is returned to the container again with the help of another pump. This method of measurement, like that described in US Pat. No. 3,714,445, is very complex both optically and mechanically, in addition both the measuring chamber and the pumps and the transport conduits of the samples must be frequently sterilized, a complexity that limits their use in routine diagnostic methods. Patent GB 2 221 986 and US patents 3,819,278 and US 4,725,148 relate to turbidometers for direct measurement of microbial growth using the same principle as those described above. They present complex optical and mechanical systems and require the sterilization of their parts between each batch of microorganisms.

On the other hand, the US Pat. No. 3,832,532 uses a conventional optical device, which includes a cuvette of spectrophotometric quality, which for the purpose of carrying out measurements using antibiotic disks includes in its design an interconnected bi-lobed reservoir, so that concluding the incubation, transfer the liquid to the other chamber in order to execute the measurement, trying to avoid the presence of the antibiotic disc to carry out said reading operation. The solution proposed in this invention presents a great complexity, both operative and technical, with the consequent economic implications. The current trends of microbiology are directed towards the search for procedures that allow the rapid identification of microorganisms (between 2-4 hours) in biological samples, and for this different strategies have been used, among which is the use of enzymatic markers. specific. According to the state of the art, most of the biological samples can not be used directly, being necessary the growth and previous isolation of the germ to identify, which constitutes a considerable loss of time that precludes a result before 24-48 hours of receiving the samples in the laboratory. Urinary tract infections are considered one of the most frequent among infectious diseases. The classical techniques for detecting bacterial infection in urine still need a plaque culture that usually requires a minimum of 24 hours of incubation to rule out all negative samples and select positive ones. It is known that only about 20% of the urines that reach the laboratory are positive and of these, 70% correspond to infection by E. coli, hence the rapid identification of E. coli allows a considerable saving of time and resources, since only 30% of positive samples would be isolated for later identification. The identification of E. coli, according to the state of the art, is carried out fundamentally through two specific enzymatic markers for this bacterium, which are the -D-glucuronidase and the Triptofanase, through the formation of Indol (Kovacks, N. Eine vereinfachte Methode zum der Nachweis der Indolbildung durch Bakterien, Z. Immunitatsforsch., 55; 311-315, 1928). 94% of all E. coli and a few Salmonellas and Shiguellas show a positive reaction to -D-glucuronidase. The formation test of Indole is positive for 99% of all E. coli, so the combination of both tests allows the unambiguous identification of this microorganism. Currently, different tests are commercialized, such as BACTIDENT-E. coli and culture media such as FLUOROCULT-MUG (both from the company MERCK DIAGNOSTICA), which are based on the previous principle. However, for the use thereof, it is necessary to previously isolate the microorganism to take an isolated colony, from which the identification is made (BACTIDENT) or the sample under analysis is inoculated in the culture medium and wait 24 hours to obtain the isolated colonies and detect the changes associated with the transformation of specific substrates (F UOROCULT-MUG). In the patent application No. WO 95/03424, a solid culture medium is reported for the simultaneous detection of coliform bacteria and / or E. coli in water or food samples, for which 24 hours of incubation are required after the inoculation of the plate with the sample to be evaluated. Similar procedures are followed in the URILINE ID Diagnostic Kit and the CPS ID culture medium, both from the company BIOMÉRIEUX, France. In both cases it is also necessary to incubate the samples in the solid medium, for 24 hours, prior to the identification of the microorganism. Patent application No. WO 80/02433 relates to a method for the identification of bacteria by combining different tests for the determination of 26 bacterial enzymes, among which are -D-glucuronidase and Tryptophanase for the identification of E. coli. In this invention bacteria must be isolated from clinical specimens before identification. Disclosure of the invention. The objective of the present invention is to provide a system that allows the detection of microbial growth early in samples from animals, plants and their secretions, in which it is required to detect growth of microorganisms by using microsamples. Said system is based on the detection of turbidimetric changes in a culture medium, produced by the growth of the microorganism, a system that includes equipment, a diagnostic kit and a method designed for this purpose. A novelty of the present technical solution is that it allows the detection of infested samples obtained directly from the specimens that produce it. Additionally, and among other applications, the present invention makes it possible to obtain the sensitivity scheme for microorganism antibiotics either from previously isolated colonies or from positive samples of urine cultures and blood cultures, in the latter case saving the time required for the processes of isolation and purification. In the particular case of urinary tract infections, the present invention allows from direct samples of urine, discriminate positive samples from negative ones, be contaminated or not with another germ, and may also include the simultaneous identification of those samples infested particularly with the bacteria E. coli. With the system of the present invention, results based on more than 1000 samples examined have shown 95% correspondence with the total count of viable cells in CLED medium, a method conventionally used for the detection of urinary system infection. In the determination of the antibiogram, the correspondence with the Bauer-Kirby method has been established in 75.8% as a predictive value for sensitive antibiotics and 85.9% for resistant antibiotics, obtaining a total sensitivity of 80.6%. The system guarantees a 90% effectiveness for the detection of sensitive antibiotics.

With the use of the present invention, it is possible to obtain useful information relative to the microbiological clinical diagnosis in a very short time, which can be used in patients to avoid the inappropriate use of antibiotics, the development of microbial resistance, long hospital stays. and fatal outcomes in serious infections. The proposed system is characterized by its rapidity, since it allows the determination of the urinary infection in 4 hours, and from positive samples it offers reliable results of the antibiogram in only 4 hours. It is also a highly accurate system that allows you to corroborate the results obtained as many times as you think. From the social point of view it is of great importance, since for those hospitalized people it makes possible the supply of antibiotics in a rational and timely manner, avoiding prolonged hospital stays. Likewise, from the ecological point of view it has a great impact since, by avoiding the inappropriate use of antibiotics, it limits the development of bacterial resistance at the same time. It is a highly flexible system that allows adapting the information of the program that it uses to the needs and demands of the user with the possibility of changing the set of antibiotics used, according to the particular needs. The system of the present invention consists of a kit, a set and a method designed for rapid microbiological diagnosis, with applications in the human and veterinary clinic. The equipment has been designed to work with a large number of samples and employs an operational program, and a very functional man-machine inferióase, which offers an audiovisual alarm to order reading and prevent operational errors. As it is a stand-alone device based on a microprocessor, it can store the results of 10 antibiograms with 14 antibiotics and offer the response on screen in 7 segments and in a printer. The measurement module of said equipment can be inserted in the free slot of the computer. The equipment of the present invention is comprised of the following devices: • A control module or microprocessor incorporated in a personal computer. • An inferred card. • A peristaltic pump. • A sensor. • A calibrator.

• A printer. • One UV lamp optionally. The personal computer must have the following properties: - IBM Compatible, 386/486, 25-66 Mhz. - RAM Memory 1Mbyte minimum. - Hard Disk: 40 Mbytes minimum, Flexible Disk Tower 3 1/2", optional - SVGA Color Monitor - Keyboard, Mouse (ouse) and Printer The peristaltic pump has been designed to circulate the samples through the sensor, which has a manual adjustable cassette that guarantees a continuous and adjustable flow, being possible its independent use or incorporated into the system.It uses a flow of 2.0 - 2.6 ml / minute and is optionally fed with 220 vac or 12 vdc, as well as its consumption The sensor is constituted by a continuous microfluidic reader, which is in charge of detecting in terms of optical transmittance, the changes of turbidity due to microbial growth in previously prepared samples from different sources, in very small sample volumes (up to 200 microliters) and in motion, that is, in a moving liquid, where the samples may or may not be interfered With the use of the present invention It is not necessary to "clean" the reader to flow during a measurement process, so that discrete or continuous samples can be measured, as long as it is not interfered with by the turbidimetric variations of the culture medium as a consequence of the sample change. However, it does detect small variations in turbidity of the medium due to microbial growth, where temperature control is not required to perform the measurement and is possible perform an indeterminate number of measurements of different samples of different origin. This sensor is also characterized by its automatic calibration and continuous flow of 2 - 2.6 ml / minute. It uses a range of conductimetric measurement of 200 - 300 micro-Siemens and an optical measurement range of 0.00 - 2.00 McFarland. The calibrator used in the present invention is based on the nephelometry technique and is adjusted by the McFarland scale. It consists of a direct light source and a photosensor and its adjustment is carried out through a program. Its function is the measurement of turbidity in liquid Mueller-Hinton medium up to 0.2 McFarland. Its power is 5 vdc and its consumption 50 ma. The UV lamp that is optionally coupled to the equipment of the present invention allows the identification of E. coli bacteria in the samples analyzed.

The program designed to operate the system object of the present invention offers a friendly and easy-to-use man-machine interface, which allows the selection of options by bar menu or function keys combined with the use of icons, as well as storage of data automatically. It also uses a structured interrogation language (SQL) for the extraction of information and in the external utilities is the 'BACKUP' that makes backup copies possible. It is equipped with an alarm system in case of violation of the entry condition, an audiovisual alarm to control the reading time of each sample, as well as other utilities for its technical maintenance. The essential distinctive features of the equipment object of the present invention are the static turbidimetric minilector and the microflow sensor that is driven by the peristaltic pump, and its coupling to a microcomputer with a program package for the acquisition, processing and creation of bases of data, which allow the generation of the necessary reports. Figure 1 shows the integral scheme of the equipment of the present invention, in a general view. As you can see, this equipment is based on a microflow turbidimetric reader (1), which is fed by a peristaltic pump (2) and fitted to a high sensitivity electronic equipment (3) that detects turbidimetric changes of (1) by means of a measurement method that allows, using a group of algorithms, detect small turbidimetric variations and proceed appropriately with the obtained data. This is formed by a turbidimetric measurement circuit (4) connected through an inferium card to a central processing unit (5). This unit receives all the keyboard commands (6) and sends the results to a screen (7). This can detect small variations of turbidity that occur in the culture medium inoculated with the sample that will be analyzed. A detailed scheme of the internal structure of the turbidimetric reader (1) is shown in Figure 2, where the detection of turbidimetric variations occurs. The measurement procedure is very simple. The nozzle (8) of the reader is introduced into the sample to be measured, which circulates therethrough with the help of the peristaltic pump (2) of figure 1, operated all the time required for the measurements. Firstly, the presence of the sample is detected when a voltage value exceeding a predetermined threshold value is obtained and after a certain time the measurement is made and so on. Because the peristaltic pump (2) of figure 1 is kept activated, there is a period of time between each measurement during which air will circulate through the turbidimetric reader (1), this time being considered as the moment in which it is performed the cleaning of the turbidimetric reader (1), that is, no additional washing process is needed to sterilize the parts that make up said reader. The measuring chamber is composed of a plastic hose (9) inserted in a glass capillary (10). The passage of light (11) is the diameter of the hole that must pass through the light from the light emitter (12) to reach the measuring chamber (9 and 10). The intensity of the light radiation transmitted through the measuring chamber (9 and 10) will depend on the degree of turbidity in the sample and is measured by a photodetector (13). The radiation emitted by the photoemitter (12) is stabilized by means of a conventional electronic control loop. Figure 3 shows the operation of the turbidimetric reader through a flow diagram. First, the presence of the turbidimetric reader is verified by means of a detection subroutine and if present, the existence of the peristaltic pump necessary for the operation of the turbidimetric reader and for the execution of the washing subroutine is verified., in addition to establishing the required workflow. The aforementioned washing subroutine is of significant importance since the parameters of the reader depend to a great extent on the cleanliness of the measuring chamber, an aspect that influences the useful life of the same. Once all working parameters have been adjusted, the turbidimetric reader will be enabled. Any subroutine that is violated will disable the operation of said reader. The fundamental application of this device is aimed at determining the sensitivity of the sample, (susceptibility of microorganisms to antibiotics), which is achieved in a period of time between 2 and 6 hours, using a diagnostic kit designed for these purposes The other device that is incorporated into the equipment of the present invention, the static turbidimetric minilector, as referred to above, makes it possible to detect changes in turbidity due to the microbial growth of the sample originating in the glass bottle containing culture medium, and which forms one of the elements of the diagnostic game of this invention. Said bottle fits exactly the reading well of said equipment. The device is adjusted to be able to use the reference bottles for direct reading of the sample, that is, no special cuvettes are required for said reading, which favors its use in routine diagnostic means. The mentioned device is used for the calibration of the inoculum used in the diagnostic set for the detection of the antibiogram, reporting the turbidity in McFarland units, in correspondence with latex standards that comply with the NCCLS standards, in the established measurement range ( 0 - 4.0 McFarland units). The diagnostic kit comprising the system of this invention is made up of a nephelometric measuring bottle of 8 ml in volume containing 4.5 ml of culture medium, which is no more than a bottle of autoclavable borosilicate glass and with a plastic lid. . Figure 4 shows the components of the diagnostic kit of the present invention, constituted by the bottle containing culture medium and the polymer, the support of the strip and the strip itself, used in the determination of the sensitivity of the sample to be analyzed. The culture medium used to follow the microbial growth is the modified Mueller-Hinton Broth OXOID medium, pH 7.4 ± 0.2. and sterile, which additionally includes a polymer. The diagnostic kit for the detection of the antibiogram of the sample to be analyzed is characterized by the use of antibiotic discs that are commercially available and that can be used forming schemes that can be varied according to any need. They are organized in non-transparent strips that include 2 free positions for the negative and positive controls, which are covered with only and half inoculated culture medium respectively, and which are used to calculate the growth index. There are also other 10 to 22 positions where antibiotic discs can be deposited. The program designed for these purposes allows the introduction of these changes in the processes of acquisition and editing, all of which gives the game a high flexibility, allowing adaptations to the most varied needs. To the above-mentioned glass bottle forming said diagnostic kit and containing the culture medium for dilution is added, according to the present invention, a polymer that can be any linear polysaccharide of structural formula CH3-CH3-Ch3-N or similar, of approximate molecular weight between 50,000 and 150,000. The incorporation of said polymer to the culture medium, at a concentration comprised in the range between 0.05 and 1%, allows to eliminate the inhibitory effect of the catabolic products that accompany the inoculum of the samples to be analyzed, obtaining higher growth rates of the bacteria involved in the infections, in a shorter period of time compared to the time required when the same culture medium is used without said polymer. This new element in the diagnostic game allows the false sensitive results obtained in the antibiotic susceptibility studies of the analyzed samples to decrease, in turn improving the correspondence with the reference method of Kirby-Bauer, 1966 (Bauer, A. W.; Kirby, W. M. M.; Sherris, J.C. and Turck, M. An. J. Clin. Pathol. 1966, 45, pages 493-496). Considering that this polymer is only metabolizable by a reduced number of microorganisms, which are not commonly found in the analyzes where the present system is applied, it is inferred that the bacterial growth promoting effect observed in its presence, is due to an inhibition of the repressing agents of said growth present in the culture medium, so that said polymer must act by absorbing and / or adsorbing the catabolites that are involuntarily incorporated next to the inoculum that is analyzed. Furthermore, it has been observed that once these catabolites are neutralized by said polymer during the process of measuring the growth of the microorganism, the bactericidal activity of the tested antibiotics becomes more specific.

One of the advantages of the present invention is that the proposed method allows to diagnose not only previously isolated and purified bacterial strains, but also direct samples of positive blood cultures, urine, etc. For the analysis of the aforementioned sample, it is first placed in the glass bottle containing the polymer and the culture medium, which is immediately determined its turbidity (tO) and this value, as well as the reading time according to the number assigned to each sample, it is fixed by the program used. Subsequently, the aforementioned bottle is incubated between 2 and 5 hours at a temperature between 35 and 37 ° C, and at the end of the incubation the system emits, according to the structured routine, an audible alarm and a sign on the screen indicating the sample that it must be read again. Those samples that register increments higher than 0.08 McFarland units, are considered as positive samples. The system of this invention allows positive samples, once detected, to determine their antibiogram very quickly. For this, an aliquot of said sample is taken and transferred to a new dilution bottle, containing fresh medium, which is then distributed in the microplate where the two controls are located (positive and negative) and from 10 to 22 antibiotic discs. . After 4 hours of incubation between 35 and 37 ° C, the plate is read, positioning the sensor at microflow in each well, following the instructions issued by the program that selects the moment of each measurement in series, eliminating the interference of reading preceding. Based on the density values obtained, the growth index (in the controls) is calculated, and the% inhibition generated for the sample by each antibiotic and according to the level obtained is assigned the criterion of sensitive, resistant or intermediate, between inhibition values comprised in the 60 and 100% range. That is, those samples with percent inhibition less than 60% can be considered resistant, those that exhibit inhibition between 60 and 80% can be considered sensitive to an intermediate level and those that are inhibited between 80 and 100% will be considered sensitive to the antibiotic in question. Each result is checked to assess if it is between the minimum and maximum admissible growth value (satisfactory antibiogram). The results obtained and the edited data of each sample are automatically passed to create the corresponding databases.

As explained above, the invention applied to the urine culture allows to work with direct samples in liquid medium, read in commercial bottles and then execute the presumptive antibiogram in positive samples, all of which is executed in a period of less than 9 hours, thus avoiding the previous steps of isolation and purification of the samples and obtaining levels of sensitivity higher than 90%. On the other hand, the system proposed here allows to face the interferences generated by the contamination of the samples, as well as by the infection caused by more than one germ. Pollution interference has been resolved by adjusting the signal in magnitude and time for the detection of internationally accepted infestation levels (> 100,000 cfu / ml), favoring the exclusion of contaminated samples not infested by counting these generally with lower bacterial levels to 1,000 cfu / ml, which allows the detection of contaminated samples only when they are also infested. In this particular case, taking into account that generally the polluting species attending the Gram are saprophytes, mostly sensitive to all antibiotics, it is evident that the contaminating strains should not interfere in the detection of the resistance scheme of the infesting strains. These also constitute distinctive features of the present invention. In relation to infections produced by more than 1 germIn practice, two particular situations can be presented. In the first, there may be a prevalence of one of the infesting germs due to a higher specific growth rate after the minimum incubation time, in which case the antibiogram would be valid. In the second, the germs grow simultaneously, in which case the antibiogram could show one or several antibiotics effective for both germs, or an absolute resistance scheme could be presented by complementation, in which situation the test could only be indicated with other antibiotics not included in the test, or later pass the necessary isolation and purification procedures for this case. This set of analyzes and immediate solutions for each particular situation, is only possible by the application of the concept of readings on direct samples, with high level of interference, at short and fixed time, aspects that characterize and distinguish the system of the present invention. On the other hand, to the report of the detection of the urinary infection and the sensitivity scheme of the infesting germ, the identification of the germ must be joined in order to have a complete report. In order to achieve this goal, the indicated bottle containing the liquid culture medium and also the polymer used for the detection of urinary infection, you can optionally add two substrates that allow to include a scheme for rapid identification of the bacteria E. coli in urine, as mentioned above, in the detection bottle itself. In the present invention, a culture medium is formulated which, when used in the proposed system, makes it possible to detect the presence of E. coli as the causal agent of the infection in urine samples tested in a period between about 9 hours. The medium proposed in the present invention contains per liter, in addition to the conventional nutritive bases contained in the Mueller Hinton culture medium, OXOID (Meat Infusion, 300 mg; Casein Hydrolyzate, 17 g and Starch, 1.5 g), substrates MU- -D-glucuronide (0.1), L-Tryptophan (1 g), as well as the polymer used as de-repressor agent (1 g). These components are solubilized in 50 mM potassium phosphate and the pH of said medium is adjusted between 7-7.5. It is then dispensed in vials with 2.5 ml and sterilized by autoclaving for 20 minutes at 121 (C. The substrates previously used are useful for the detection of enzymatic activities -D-glucuronidase and Tryptophanase produced by E. coli, by means of the formation of Indol, so that to said medium after the bacterial growth, an auxiliary reagent for the development is added (modified Kovacks reagent) composed of para-dimethylaminobenzaldehyde (2 g), Ethanol and concentrated Hydrochloric Acid (20 ml) The product of the interaction of the enzymes with their substrates is detected in a first step, exposing the vials where an increase of the turbidity is detected (positive) to a light source U.V. (attachment that may be included in the equipment of the invention optionally for this purpose), for the detection of fluorescence generated by the release of 4-methylumbelliferone. In a second phase, the formation of Indol is checked in the vial itself by developing it with said modified Kovacks reagent.

EXAMPLES OF EMBODIMENT OF THE INVENTION: Example 1. Start-up of the Equipment: The equipment is turned on 15 minutes before starting the readings. When activating the computer, the self-execution program leads directly to the program in charge of executing the entire process, which will initially check the existence and integrity of the database and the operating status of each of the components of the measurement module: sensor, the inoculator calibrator and the peristaltic pump. It will also check the existence of the electronic protection key that attaches to the parallel port of the computer. The program will inform about any error that is detected in any of the elements that make up the system and will disable the options related to it. If the database does not exist, it will be created if the user so decides. Example 2. Preparation for the determination of urine cultures. The urine culture is a well-known test used to detect if a urine sample is infected or not. The procedure starts with a first reading in an initial time (TOh) and continues with a second reading, usually four hours after the sample has been incubated (TFh). The determination is carried out photometrically using the inocula calibrator. The following steps illustrate the procedure to follow: 1. Take a bottle with 4.5 ml of sterile culture medium to which 500 μl of the urine sample is inoculated. 2. The initial McFarland turbidity is measured using the inoculum calibrator (TOh). 3. The sample is incubated at 37 ° C for 4 hours, and the equipment warns, by means of an audible alarm when this time has elapsed for each sample. 4. A next or last reading of the inoculated bottle is made after its incubation to check if the sample is positive, negative or doubtful (TFh). In the case of samples reported as "doubtful", the incubation should be prolonged 1 hour more, to rule out its positivity. In cases of negative samples from patients with suspected renal infections, it is advisable to prolong the incubation up to 5 hours (T5). Once the reading is done, the cases can be edited if desired and stored in the system database. If the automatic printing mode is activated and the printer is ready to print, the cases will be automatically printed. Samples that have been diagnosed as positive can be processed for the determination of the antibiogram (directly). Example3. Preparation for the antibiogram. For the antibiogram 12 or 24 wells strips are used. The first 2 wells are reserved for the positive control (C +) and negative control (C-) respectively and in the remaining wells are the antibiotic discs.

Inoculation of the strip is done by distributing the test sample in the first well (C +) and the wells containing the antibiotic discs. The second well (C-) must be filled with sterile medium. Example 4. Preparation of the inoculum. • From a pure strain. If the inoculum is prepared from a pure strain, the following steps should be carried out: 1. Take 3 to 4 colonies of a fresh culture (18-24 h) and inoculate 4.5 ml of sterile Muller Hinton broth containing a linear polymer of molecular weight 50,000, until reaching a concentration of cells equivalent to 0.5 of the McFarland scale in the mininephelometer. Option presented in the Main Menu (McFarland). 2. From this cell concentration, 150 μl is taken and added to 4.5 ml of another sterile medium, and stirred to homogenize. 3. From this dilution, 200 μl (0.2 ml) is distributed in the strip for the antibiogram, in the wells corresponding to the positive control (C +) and in the wells containing the antibiotic discs. 4. In the second well destined for the negative control (C-), 200 μl (0.2 ml) of sterile medium is dispensed. 5. Upon completion, the strip is sealed again and incubated at 37 ° C for 4 hours. 6. Once the incubation time has elapsed, the strip is removed from the incubator and 10 minutes are waited until the room temperature is reached and the reading begins. It is recommended to shake the strip manually or mechanically before reading to homogenize the contents of the wells. 7. The strip is uncovered and the reading is started. It is required to read the antibiogram that the difference between (C +) and (C-) reaches a certain value defined by the Minimum Admissible Growth. If in the presence of values close to the differentials established by this parameter, each of the tested antibiotics is detected, homogeneously low levels of inhibition (Antibiogram not reliable), the antibiogram must be repeated. In these cases it is recommended to decrease until reaching 30% the volume of inoculum used. If the inoculum is prepared from a positive urine culture, to perform the antibiogram, it is necessary to pre-dilute the sample until reaching 0.5 of the McFarland scale and once the desired turbidity is reached, the process is continued from point 2 described - previously to obtain the inoculum from a pure strain. In cases in which the urine culture is detected as positive in 4 hours and has values below 0.5 of the McFarland scale, it must be incubated until it reaches that value or it can be fluted in a medium of its choice to be worked the next day, that this may be due to: 1. Low load of non-infesting microorganism. 2. Patient under the action of antimicrobial therapy. When the inoculum is prepared from positive blood cultures (from 18 to 24 hours of incubation), the following preliminary steps should be carried out: - Extraction of the upper blood culture phase (supernatant) 200 μl and add them in 4.5 ml of Mueller medium -Hinton + Pol-10. Read turbidity in the mininephelometer in the McFarland option of the Main Menu and monitor until it reaches values between 0.5-0.7 of said scale. - The process is continued from point 2 described above to obtain the inoculum from a pure strain.

Example 5. Use of the programs. The invention provides a package of fully interactive programs that allow the testing of antibiograms, urocultures, store and process the results in an environment that is easy to operate and does not require previous experience in the management of computers. Each test is treated in independent menus that group the fundamental operations performed in each of them. To access any of the system options: 1. Press the key corresponding to the underlined letter inside the desired icon. 2. Place the mouse cursor (mouse) over the selected icon and press the left button. To abandon the chosen operation, press the "ESC" key or activate with the mouse the option "Return" which will lead to the previous screen. After completing the initiation of the system the program will show the menu of the MAIN SCREEN that constitutes the starting point for all the operations that you wish to carry out with it. As you can see, the main screen is composed of three zones or panels: - A central panel that displays the icons that activate the main options of the program. - A top panel, where information is provided on the system version, the current date and time, the menu in which you are working and an indication on the action level of the screen that is displayed. In the upper right corner, the option "Exit" is offered through which you can leave the program at any point as long as it is not in the middle of a measurement or some critical operation. It is advisable to always use this option and NEVER SHUT OFF THE COMPUTER without first closing the program. Ignoring this recommendation can lead to problems in the configuration of the programs. The option "Ref. P." that appears at the same end of the screen, offers the possibility to activate the screen refresher at will when it is not working, in order to protect it from the marks that can cause a fixed image for a very long time. This option is also activated automatically when the idle system is detected during a certain time. To return from this option, just move the mouse (mouse) or simply press any key. - The lower panel reports on the status of urine culture tests. In case there are urine cultures pending to be read, we will inform how many, as well as the date and time of the first in the list ordered by next reading. It is also indicated if the sound signal that the system emits indicating that the time has come to read a certain urine culture, is active or not. Main menu . The main menu consists of a set of icons that contain drawings related to the operation performed when they are activated. A black triangle pointing to the right in the options "URO and ATB "is an indicator that warns that activating one of them will show a submenu with additional options.Nine icons define the options available within the main menu, from where all the actions of the system are controlled.These are: URO: Activates the menu of urine culture icons ATB: Activates the antibiogram icons menu McFarland: Allows measuring the content of the bottle in the calibration unit (McFarland scale) Database: Accesses the antibiogram or urine culture database. : To change the system configuration Information: Displays administrative information of the system.

DIRAMIC: Provides the address of the system producer. Help: Displays information about the current option. Exit: Leave the system. Uroculture menu. When selecting the URO icon from the Main Menu, a new menu will be available, specific for the treatment of urocultures, which contains the operations to be developed during these tests. Within the menu of UROCULTIVOS the following operations can be carried out: Description of the options of the Uroculture Menu: • Read Uros: Procedure for the reading of urine cultures.

When activating the "Read Uros" option, a list of urine cultures that are pending to be read is displayed, showing the consecutive number of the case, the time between readings and the date and time of the next reading. If the list is empty, there are no pending cases to be read. To select a specific case of the list you can press the left mouse button (mouse) twice relatively quickly on it or simply press the space bar when the mouse cursor is positioned on it. The selected ones will appear with a mark next to them and most of the operations will be carried out only by them.

By selecting the Read Uros option, the following possibilities will be available: • New Uros: Allows you to read the time Oh of the uroculture cases. If the inocula calibrator has not been compensated, the program will ask that a bottle be inserted with means to proceed with the compensation before adding the new cases. Activating this option will show a screen where you can proceed with the reading, edit the case (Edit) or change the time that will pass until the next reading is executed (TO-TFh) BEFORE reading it • Re-read: Allows you to return to read the time Oh of a sample. This option is useful when you have made an error in reading one of the cases and you want to rectify it. Activating this option will show a screen with the same possibilities as the "New Uros" option plus the option to continue to the next case (Next) without modifying the one being processed. • Edit: With this option you can fill in the fields corresponding to the general patient data, those of interest to the laboratory, etc. To store the information that has been edited, press the left mouse button (mouse) on the desired option. Optionally you can use the RETURN (or ENTER) keys to accept the information and ESC to cancel the editing in process. The user can freely edit all the fields with the exception of those corresponding to Sample, Microorganism and Area, which are differentiated by a representative indicator of a selection process against a list of options. By placing the mouse cursor over the corresponding fields and pressing the left button, a selection list opens offering the possibility of performing an intelligent search within it by pressing the first letters of the word that constitutes the desired objective. You can add or delete specific names within the preset lists using the functions provided in this window. This option is used to maintain a standardized listing of the data that is inserted in the database. If any element of the list that is displayed is modified, all the existing cases in the database will be updated accordingly. To jump from one field to another within the editing process, press the TAB key or simply press the left mouse button (mouse) on the desired field. If you are editing several cases, the "Next" option will appear with which the current case edition is ABANDONED without storing the changes and the next case is automatically edited. In a different way, if the "Accept" option is activated, the case data will be stored on the screen before moving on to the next one. • TO-TFh: It allows configuring the time that will pass between the first and next reading. Note: If the time for which you want to configure the case has already elapsed, the program will warn of this and show an error message. This option will be executed for all selected cases whenever possible. • Cancel: Allows you to cancel or delete unwanted pending cases. This option will be executed for all selected cases. • Read TFh: It allows to execute the last reading of the urine culture in order to obtain the result of the examination. When activating this option a screen will appear with the general data of the case and its classification according to the level of infection of the same in Positive, Negative or Doubtful. This option will also be executed for the selected cases. "Next" in this case does not read or classify the case and shows the following which is useful when you want to increase the time between readings and wait more to see its evolution • Selection: Allows you to mark or unmark all cases in the urine culture list . • Sort: Allows you to sort the list by consecutive number of samples or by next reading. This option is useful when you want to sort the cases by hour in which they should be read that will not always coincide with the order of the consecutive number. • Print: Allows you to print the case to the printer or to a file in ASCII format. When this command is activated, a printing dialog will be presented where the following parameters can be configured for the output of the information: • Printer: Allows you to select a type of printer compatible with the one that is attached to your computer. • Epson: For dot matrix printers. • HP LaserJet: For laser printers. • Adobe PostScript: For printers with PostScript format. • ASCII: To print to an ASCII file. This possibility, very useful when you want to export data from the program, allows you to specify the name and location of the text file where the data will be stored.

• Cancel: Return to the previous option. • Copies: Number of copies to print. • Lines / Pag. : Number of lines of text on each page. • Print to: Allows you to select the output port of the computer where the printer is installed, generally LPT1, LPT2, LPT3 or LPT4. • Return: Allows you to return to the previous option. Within the print dialog, the following commands are available: • Accept: Start printing. Activating this option will show an information message about printing with an option to cancel it if desired. • Cancel: Cancels the operation. • Return: Transfer the control to the UROCULATIVE MENU Report: Provide a concise report on the results of the urine culture test on the date selected by the user, allowing the user to print it. McFarland: This operation allows to monitor the microbial growth, expressed in McFarland units according to the NCCLS standard, of a sample referred to a previously established target (sterile non-inoculated culture medium). The samples are placed in the measuring well of the calibrator previously compensated and the information provided on the screen is read directly. For this purpose, an approximate graphic scale and the digital value of the reading are shown. The second decimal figure of the digital values that are presented constitute, fundamentally, a measure of the trend of the sample analyzed. As a standard of work, it must be ensured that the outer surface of the bottles that are placed in the well of the inoculator calibrator is clean and free of scratches. The bottles should be marked conveniently in order to repeat their position in each measurement, taking care that the mark made is outside the measuring well. Following this practice, adequate accuracy will be achieved. Two commands are available for calibrator operation: • Compensate: Set the target or reference for the determination of turbidity or McFarland value of the samples. This option is particularly useful if a different batch of culture medium will be used after the Calibrator was compensated. The process is achieved by positioning a bottle containing uninoculated culture medium in the measuring well of the McFarland calibrator. • Return: Return to the previous option. Database: Through this option you can print and visualize the results of antibiogram and urine culture cases performed with the system. In the same way you can modify the general data and execute searches under certain criteria which facilitate the study of the cases carried out. As information, the current case that is being visualized and the total number of cases that exist in the base are shown in the upper area of the window (eg Case: 1 of 100). The way in which the data is displayed is also shown, that is, "All cases" means that all cases of the base or "Search" are shown, which are the cases that respond to certain conditions established during a specific search. The word "Visualizing" informs if what is presented on the screen are the "General Data" or the "Results" of the exam. The options available for managing the database are the following: • First: It allows viewing the first case of the database.

• Previous: Allows visualizing the previous case to the one that is being viewed. • Next: Allows visualizing the case that follows the one that is being viewed. • Last: Allows viewing the last case of the database. • Go to the case: It allows to visualize a determined case according to its number in the database. • Data / Results: Allows viewing of general data or test results. • Edit: This procedure has already been described in the section Read uros. • All / Search: Allows you to view all cases in the database or execute a query or search: Within this procedure, you can have the following options: • Search: Allows you to execute the search or query according to the formulated expression . • New: Allows you to formulate a new search expression. • Return: Allows you to return to the previous option. Some of the available "Search" criteria are explained below.

• Contains: Allows searching without taking into account if the data was written in uppercase or lowercase. It also allows you to search for words or syllables that are found in a specific word or phrase (eg "infec" in "Severe Infection"). • That does not contain: It works like the previous one with the difference that excludes cases that meet this condition. • That is in: Allows you to create a list with the desired options. This criterion is used when you want to search for several microorganisms, etc. Activating this option will show a list where elements can be added or deleted. • That is not in: It works like the previous one with the difference that excludes cases that meet this condition. • Do not process: It allows deleting the data of the formulated search expression. • Ignore: Ignores the selected field during the search process. • "Equal to" or "Different from": Compares identical words, taking into account if their writing differs by the use of lowercase or uppercase. To do a search that does not consider whether the words were written in one way or another, you must use the criteria "containing" or "not containing" as appropriate. • Print: Allows you to print the case to the printer or to a file in ASCII format. • Return: Return to the UROCULS MENU. Options: Allows you to configure the following possibilities: • Automatic printing after the sensitivity measurement. • Activate or deactivate the sound signal that indicates the moment in which a certain urine culture is ready to be read. • Decide on the use or not of colors in the presentation of the report. • Set the time that will mediate between the two readings of a urine culture (normally the program will establish an interval of 4:00 hours). Compensate: Adjustment of the McFarland gauge. It allows to establish the level of reference for McFarland = 0. This process is carried out using a bottle with culture medium without inoculation.

Consecutive: It allows configuring the consecutive number for new cases of urocultures whenever possible and does not cause conflicts with the cases already read. This option is convenient when you want the consecutive counter of urine cultures to start with a different number of 1. For example, start at 50, 100, etc. Note: The program considers the consecutive number as the "consecutive of the day". When detecting a change in the date, it will try to take it back to 1. Help: It shows information to the user about the system and each one of its operations and options.

Return: Transfer the control to the Main Menu. ATB antibiogram menu: Antibiogram menu By selecting the ATB icon from the main menu, the control is transferred to the operations to be developed during this test. There are twelve and are described below: Read ATB: It allows the reading of an antibiogram that will determine the antibiotic susceptibility scheme of a determined sample. Before starting the reading, the built-in control system checks systematically and automatically if the sensor has been washed and compensated and if the peristaltic pump is working correctly. This procedure is transparent to the user as long as no difficulties are detected. Otherwise, the user will be informed of the problems detected during self-check. When the first reading of an ATB of the day is executed, the system will proceed to automatically compensate the point of operation of the sensor for which, in a totally interactive way, it will guide the user during this process. Once the adjustment is complete, the system will report the "Compensation Constant", which is a numerical value that is normally below 2.00. However, values above 2.00 but less than 2.50 will be accepted and the user will be warned about the convenience of installing a new sensor. Once all the technical requirements have been met, the editing screen will be presented where the general data of the case to be processed will be filled.

The template for the edition of patient data is composed of the following fields: - Clinical History - Name - 1st Surname - 2nd Surname - Age - Sex - Date of examination - Sample * - Description - Microorganism * - Area * - Doctor - Nosocomial - Deceased At this moment two commands will be available: • Return: Return to the ATB Menu. • Accept: Selecting this option will lead the user to start the measurement, where each antibiogram is identified by its case number. In the Title Bar of the measurement window, the indication of inserting or removing the sensor from the sample will appear as appropriate. Simultaneously, a low frequency audio tone will indicate that the system waits for the beginning of the measurement of one well while another audio tone, of sharper frequency, will indicate that the measurement corresponding to that well has finished and that it should be removed. the sensor. This procedure will be repeated for each well of the strip. The set of antibiotics in use and the case number will be displayed in this window. During the measurement, the system constantly monitors any possible operating error, offering instructions to resolve any conflict that may occur. The following diagram illustrates the sequence of each reading cycle: READING CYCLE Message: Insert the sensor + Bass Sound = > Sensor inside the well + Value of the reading Message: Extract the sensor + Acute sound = > Remove the sensor from the well and move to the next one. During the measurement, the available options are the following: • Previous well: Jumps back. It allows to repeat the measurement of the previous well to the one being measured. This function is useful when an inappropriate value has been detected and you want to corroborate the measurement.

• Next well: Jump to the next well. It allows to continue with the measurement of the next well to which it is being measured and to abandon the reading of the current one. • Return: Cancels the antibiogram measurement process and returns to the previous option. When a measurement is normally concluded, the results of the antibiogram will be shown, where the tolerance of the germ to the action of the antibiotics confronting it is classified. For each antibiotic, one of three possible effects will be defined: SENSITIVE, INTERMEDIATE or RESISTANT as well as the percentage of inhibition of growth provided by each one together with the values that resulted from the readings of each well. The results are presented in units normalized with respect to the value of the Negative Control. Another parameter provided is the Minimum Admissible Growth which allows to analyze if the growth rate of the microorganism was sufficient to guarantee the validity of the antibiogram. Additionally, the built-in control system provides a classification of antibiogram validation as a measure of the reliability of the results obtained. This classification can be: Satisfactory Antibiogram: The measurement made is legal and the results are reliable. Antibiogram Not Reliable: It has not been reached Minimum Admissible Growth. - Invalid Antibiogram: The mathematical analysis of the Inhibit factor yields a set of abnormal data. URO - ATB: It allows linking an Antibiogram to an existing urine culture, inheriting the patient data supplied during the edition of the URO. Usually this option is used when you want to perform an antibiogram to a urine sample previously detected as positive. To select the case to which the test will be done, only the consecutive number and the date of the examination are required. This type of case will be identified with the word URO-ATB and the case number in the Urocultivo database. McFarland: Activates the McFarland calibrator to monitor microbial growth. Database : It allows to display and process the stored data of antibiogram cases. The procedure is similar to that described in UROCULATIVE MENU (Database). Options: In a similar way to the existing option within UROCULTIVES MENU, it is possible to choose between the following operation variants: - Enable the automatic printing of the reports of the antibiogram cases immediately after finishing the measurement. - Activate or deactivate the sound signal that indicates the beginning and end of the measurements of each well. - Decide on the use of colors in the presentation of the report. - Set the time that will mediate between the two readings of a urine culture (normally the program will establish an interval of 4:00 hours). Antibiotics: Allows to put into systematic use a selected set of antibiotics, as well as edit new games, modify and / or delete existing ones. The antibiotic scheme defined by the user has to play exactly with the antibiotic discs that exist on the plate or strip. Selecting the option "Antibiotics" will offer the following possibilities: • New: It allows to create a scheme for a set of antibiotics. After the selection, the user will ask for the number of strips and the number of wells per strip in order to build the corresponding scheme (Normally 2 x 8). Then, a window will be displayed with the aim of filling the joint name and selecting, from the list provided, each antibiotic that will be used. • View: Allows you to view or print a specific set of antibiotics. • Modify: Change the distribution or eliminate antibiotics from a specific set. • Eliminate: Eliminate a complete set of antibiotics. • Update: Predetermines the set of antibiotics that will be used in the next antibiograms. To change the current placement, the desired set should be selected and the "Update" option should be activated after the selection is made. • Select all: Select all existing sets of antibiotics. • Do not select: Leave without selecting each set of antibiotics previously chosen. • Return: Return to the previous option.

Stability: This procedure is used to test the functioning of the sensor. Statistical parameters such as mean value of the measurements, standard deviation and variance coefficient of a set of 16 identical samples are calculated. The results are stored in a database from where they can be retrieved to view or print, to user selection. For its execution, use distilled water for each of the 16 measurements. The expected values for these statistical parameters are: 2000 < Average Value: < 3000 Standard Deviation < 20 Variation Coefficient < 2.00 Cleaning the Sensor: It allows executing a procedure to carry out the cleaning of the sensor at the operator's discretion, in which, unlike the one systematically guided during the normal operation of the equipment, the different times that make up the process can be programmed. This cleaning consists of 3 basic stages: 1. A first time during which biological detergent is circulated to the flow sensor using the peristaltic pump. 2. A second rinse time with distilled water 3. One last drying step of the sensor. Calibrate: Execute the sensor calibration process. Calibration is defined as the procedure by which the response levels of each sensor are determined against an established bacterial growth and the update of this parameter as the time of use of the same elapses. The calibration must be carried out whenever a new sensor is installed. Additionally, the system is programmed to recommend, with certain regularity, the convenience of carrying out this operation for updating purposes. This process does not constitute a daily routine, so its execution is recommended whenever the automatic suggestion of the system appears in order to guarantee the level of sensitivity required for a good operation. To perform the calibration of the sensor, have one bottle with sterile culture medium (C-) and another with inoculated culture medium (C +) preferably with Staphylococcus aureus, grown to a level of 0.5 McFarland. Use the McFarland gauge to determine the proper growth.

Use a second vial containing sterilized culture medium in order to obtain a McFarland 0 (C-) index. Select "Calibrate" from the ATB Menu and follow the interactive procedure in order to execute three alternative readings of one (C +) followed by one (C-). The system will calculate the average value of the three measurements and the final result will be stored after selecting the "Accept" option. Check flow: This is an auxiliary resource to facilitate the checking of the flow rate and the continuity of liquid supplied by the peristaltic pump. The discontinuity of the sample flow can lead to measurement errors. The flow is measured using a 10 ml graduated cylinder and a container with distilled water. To do this, insert the inlet end and the outlet end of the sensor hose (both) into the water container and activate the PROCEDURE command. From this moment, the program will guide you through the process. Finally, verify that the flow of the peristaltic pump is in an environment of 2.4 ml / min. If the proper flow rate is not reached, check the condition of the Silicone hose, especially the portion inside the cassette of the peristaltic pump.

Also note the tension of the same and the adjustment of the cassette. If the hose is excessively damaged or collapsed, install a new one. Help: Provides information to the user about the system and each of its operations and options. Return: Return to the Main Menu. Example 6: Results of clinical studies conducted in Cuba: A total of 567 urine samples were analyzed for the presence of significant numbers of uropathogens using the system of the present invention, and comparing the results with the reference method CLED (semi-quantitative method of plate culture of Clarigde). Of the total analyzed, 126 samples were positive by CLED, while 108 were using the present system, but only in 4 hours, that is, while the CLED method gives the results between 24 and 48 hours after inoculation of the medium of culture, the present system was 86.1% effective in detecting the positive samples at only 4 hours after inoculation of the sample. Of the 441 negative samples found by CLED, the present system was able to detect 440 negatives, also in a 4-hour period, for an effectiveness rate of 99.8%. The general correspondence of the present system with respect to the traditional CLED method was 89.1%. Example 7: Results of clinical studies conducted in Canada: In another study conducted in Canada, 1,016 urine samples were prospectively investigated. The results obtained with the present system were compared with the CLED method for the detection of bacteriuria. For routine culture, 0.001 ml of urine were seeded on CLED agar plates, using a calibrated pipette. This method detects >; 1000 colony forming units / ml (cfu / ml). Turbidity readings were made on time 0, 2, 3, 4 and 5 hours after inoculation. In both cases, 184 positive samples were detected (> 0.4 McFarland units). The distribution of time and the correlation with a routine culture of these samples are shown in Table 1.

TABLE 1.

As can be seen, the total sensitivity of the system of the present invention was 86.4%. Its specificity, that is, the ability to detect true negative samples as defined by the routine culture was 98.5%.

BRIEF DESCRIPTION OF THE DRAWINGS: Figure 1 shows the integral scheme of the equipment of the present invention, in a general view. As you can see, this equipment is based on a turbidimetric microflow reader (1), which is fed by a peristaltic pump (2) and fitted to a high sensitivity electronic equipment (3) that detects turbidimetric changes of said reader ( 1) by means of a measurement method that allows, using a group of algorithms, detect small turbidimetric variations and proceed appropriately with the obtained data. This is formed by a turbidimetric measurement circuit (4) connected through an inferium card to a central processing unit (5). This unit receives all the keyboard commands (6) and sends the results to a screen (7). This can detect small variations of turbidity that occur in the culture medium inoculated with the sample that will be analyzed. Figure 2 represents a detailed diagram of the internal structure of the turbidimetric reader (1) of Figure 1. The nozzle of the reader (8) is introduced into the sample to be analyzed, which then circulates through it with the help of the pump peristaltic (2) of Figure 1. The measuring chamber is composed of a plastic hose (9) inserted in a glass capillary (10). The passage of light (11) the diameter of the hole through which the light coming from the light emitter (12) must pass through to reach the measuring chamber (9 and 10). The intensity of the light radiation, transmitted through the measuring chamber (9 and 10) will depend on the degree of turbidity of the sample and will be measured by the photodetector (13). The radiation produced by the photoemitter (12) is stabilized by means of an electronic loop of conventional automatic control. 1 Figure 3 shows the turbidimetric reader working, by means of a flow chart. First, the presence of the turbidimetric reader is verified by means of the detection subroutine and if it is present, then the existence of the peristaltic pump is checked, which is necessary for the operation of the turbidimetric reader and for the execution of the subroutine cleaning, in addition to stabilizing the work flow. Once all the parameters have been regulated, the turbidimetric reader will be ready to work. Any subroutine that is violated will disable the operation of the turbidimetric reader. Figure 4 shows the components of the diagnostic kit of the present invention, constituted by a bottle containing culture medium and the polymer, the support of the strip and the strip itself, used in the determination of the antibabiogram of the sample to be analyzed. Figure 5 shows two screens related to the program that follows the main procedures for the execution of the method object of the invention. Main Menu Screen (5A): It constitutes the starting point for all the operations that you wish to carry out with it.

Nine icons define the options available within the main menu, from where all the actions of the system are controlled. These are: URO: Activates the uroculture icon menu. ATB: Activates the antibiogram icons menu. McFarland: It allows to measure the content of the bottle in the calibration unit (McFarland scale). Database: Allows access to the database of antibiograms or urocultures. Options: To change the system configuration. Information: Shows administrative information of the system. DIRAMIC: Provides the address of the system producer.

Help: Displays information about the current option. Exit: Leave the system.

Uroculture Menu Screen (5B): Represents a specific menu for the particular treatment of urocultures which contains the operations to be developed during these tests, as a representation of the main options offered by the Main Menu.

Claims (10)

1. NOVELTY OF THE INVENTION Having described the present invention, it is considered as a novelty and, therefore, is claimed as property contained in the following: CLAIMS: 1. Microbiological diagnostic equipment characterized because it has two fundamental devices: a static turbidimetric mini-reactor and a microflow sensor, operated by a peristaltic pump and coupled to a microcomputer by means of an interface card, and optionally an attachment constituted by a UV lamp for the identification of E. coli in the samples analyzed.
2. 2. Equipment according to claim 1, characterized in that the turbidimetric minilector is constituted by a small measuring chamber composed of a plastic hose (9) inserted in a glass capillary (10) which is illuminated by means of a light passage (11). ), orifice that allows the light coming from a light emitter (12) to reach the sample that is analyzed and that circulates through the measuring chamber (9 and 10) with the help of a peristaltic pump (2).
3. 3. Equipment according to claim 1, characterized in that the microfluidic sensor is constituted by a photodetector (13) that measures the degree of turbidity existing in the sample, depending on the intensity of the luminous radiation reaching it after passing through the camera. of measurement (9 and 10).
4. 4. Equipment according to claims 1 to 3 characterized in that the set consisting of the turbidimetric minilector and the microflow sensor is coupled to a high sensitivity electronic equipment (3) that includes an electronic loop of automatic control in charge of stabilizing the radiation emitted by the photoemitter (12) and detects by means of the photodetector (13) the turbidity changes that occur in the sample circulating through the measurement chamber (9 and 10) of the turbidimetric reader (1), as well as it is connected to a central processing unit (5) that receives the commands of a keyboard (6) and delivers the results on a monitor (7).
5. 5. Game for the microbiological diagnosis characterized in that it uses a non-transparent strip that includes 2 free positions for the positive and negative controls and also others from 10 to 22 positions where antibiotic discs can be deposited at option; a glass bottle containing culture medium and a polymer, said bottle optionally containing for the identification of E. coli enzymatic substrates and additives that optimize said enzymatic activity, as well as an optional additional bottle containing a reagent for the development of said activity enzymatic Diagnostic kit according to claim 5, characterized in that the culture medium used is Muller Hinton liquid medium (OXOID) modified. Diagnostic kit according to claim 5, characterized in that the polymer added to the culture medium is a linear polysaccharide of structural formula CH3-CH3-Ch3-N, with an approximate molecular weight between 50,000 and 150,000, which is added to said medium at a concentration comprised in the range between 0.05 and 1%. 8. Diagnostic kit according to claim 5, characterized in that for the identification of E. coli, the vial containing the culture medium and the polymer additionally contains the substrates MU- (-D-glucuronide and L-Tryptophan solubilized in Phosphate Potassium 50 mM, pH between 7-7.5 9. Diagnostic set according to claim 5 characterized the developer reagent employed is the modified Kovacks reagent, consisting of 2 g of para-dimethylaminobenzaldehyde diluted in ethanol and 20 ml of concentrated hydrochloric acid. Method for microbiological diagnosis characterized in that it comprises the following steps: a) An aliquot of the obtained sample is placed directly from the source that contains it in the glass bottle containing the culture medium, the polymer and optionally the selected substrates and the additives necessary for the identification, determining the turbidity of said vial with the previous content on time 0; b) The bottle is incubated between 2 and 6 hours at a temperature between 35 and 37 ° C; c) The growth index between time 0 and the selected incubation time is determined and the positive samples are discriminated from the negative ones depending on the increase of the turbidity, being positive those with increments higher than 0.08 McFarland units, of which take aliquots for further steps; d) To perform the antibiogram, take an aliquot of the positive sample determined in step c) and transfer it to a new dilution bottle containing fresh medium, which is then dispensed at a rate of 200 1 in each test well. of the strip, and its incubation is carried out between 3 and 4 hours at a temperature between 35 and 37 ° C; e) The plate is read by positioning the sensor at microflow in each well, following the instructions issued by the program designed for that purpose; f) From the density values obtained, the growth rate in the controls is calculated, and the% inhibition generated for the sample by each antibiotic, and according to the level obtained, the sensitivity criterion is assigned in the range between 60 and 100% inhibition, after checking the inclusion of the result between the minimum and maximum admissible growth value for the microorganism in question; g) The results obtained from the previous step and the edited data of each sample are automatically passed to create the corresponding databases for the establishment of the antibiogram; h) For the identification of positive samples infested by E. coli, an aliquot of the bottle grown in step c) is subjected to a light source U.V. , the fluorescence generated therein being detected by the release of 4-methylumbelliferone, followed by the formation of Indol by development with modified Kovacks reagent; i) In the event that the sample has not been identified as E. coli in step h), the aliquot previously obtained in step c) is subjected to the traditional identification procedures.