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WO2000052196A1 - Procede de detection de microbes resistant aux agents antimicrobiens - Google Patents

Procede de detection de microbes resistant aux agents antimicrobiens Download PDF

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Publication number
WO2000052196A1
WO2000052196A1 PCT/DK2000/000076 DK0000076W WO0052196A1 WO 2000052196 A1 WO2000052196 A1 WO 2000052196A1 DK 0000076 W DK0000076 W DK 0000076W WO 0052196 A1 WO0052196 A1 WO 0052196A1
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Prior art keywords
species
sample
resistant
cells
group
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English (en)
Inventor
Birgitte Fauerholm Christensen
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N Foss Electric AS
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N Foss Electric AS
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Priority to AU26589/00A priority Critical patent/AU2658900A/en
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Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/045—Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/10—Enterobacteria
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/14—Streptococcus; Staphylococcus
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a method whereby it is possible to screen samples of interest for the presence or absence of microbial cells that are resistant to one or more antimicrobial compound to which such cells are normally susceptible.
  • the method is particularly useful in the screening of larger numbers of samples for the presence of cells including cells of pathogenic species that are resistant to a range of antimicrobial compounds such as antibiotics.
  • antimicrobial agents in particular antibiotic compounds
  • antibiotic compounds have been the most important means of controlling infectious diseases in animals and humans.
  • the usefulness of these highly valuable medicaments has been endangered by the occurrence of pathogenic strains that have developed or acquired resistance to one or more antibiotics.
  • the implication hereof is the choice of antibiotic becomes complicated, as the medical doctor or the veterinarian who is responsible for treating the infectious diseases needs information on the susceptibility of the particular pathogenic strain infecting his or her patient to make an appropriate choice of treatment.
  • the pathogenic strain is resistant to all of the available antimicrobial medicaments or it is only susceptible to compounds that in addition to their antimicrobial effect have serious adverse effects on the patient to be treated.
  • the widespread occurrence of antibiotic resistant microorgansms in animals and human beings is primarily the result of selection of spontaneously occurring mutants that have acquired a higher tolerance to the antibiotic than the wild type parent strain.
  • the selection of such resistant strains is caused by the use of antibiotics in human and veterinary medicine for treating and preventing infectious diseases and as animal growth promoters in animal husbandry.
  • the dissemination of resistant microorganisms has been accelerated by the occurrence of transmissible plasmids carrying resistance genes, which plasmids can be transferred within a species or among species. These resistance gene-carrying plasmids are frequently carriers of multiresistance, i.e.
  • the host cells By acquiring such a plasmid the host cells become multiresistant towards a range of the most commonly used antibiotics such as tetracyclines, antibiotics of the ⁇ -lactam group, aminoglycosides, amphenicols or sulphonamides.
  • antibiotics such as tetracyclines, antibiotics of the ⁇ -lactam group, aminoglycosides, amphenicols or sulphonamides.
  • bacteria may also carry one or more resistance genes in the chromosome.
  • such multiresistant cells may be resistant to heavy metal ions such as mercury ions and one or more preserving agent(s) or disinfectant(s) .
  • Multiresistant strains occur frequently among pathogenic or potentially pathogenic microorganisms such as e.g. Staphy/ococcus strains, E.coli strains and strains of Salmonella species and serotypes.
  • pathogenic or potentially pathogenic microorganisms such as e.g. Staphy/ococcus strains, E.coli strains and strains of Salmonella species and serotypes.
  • Such bacterial strains can be carried not only by animals and humans having clinical signs of infection but also by apparently healthy subjects including animals that are slaughtered and used to provide meat and meat products.
  • Such healthy animals carrying multiresistant bacteria constitute a significant source from which such bacteria can be transmitted to the human population either by direct contact or via ingestion of contaminated meat and offal or other products.
  • an appropriate number of cells of the isolated strain to be tested is inoculated into a liquid medium containing a serial dilution of the antibiotic.
  • the sensitivity of the strain is determined as the minimum inhibitory concentration (MIC) of the particular antibiotic.
  • the present invention provides a method of detecting in a sample the presence or absence of microbial cells of at least one pre-determined species or group of species, said microbial cells being resistant to at least one antimicrobial compond at a level to which said cells are normally susceptible, the method comprising (i) a selective enrichment step comprising subjecting the sample to at least one first enrichment step in a medium and under conditions that will permit selective enrichment of both resistant and non-resistant cells of the pre-selected species or groups of species, followed by subjecting the thus enriched sample to at least one second enrichment step in a medium and under conditions that will permit selective enrichment of resistant cells of the pre-selected species or group of species, and (iii a step of detecting in the medium that permits selective enrichment of resistant cells the presence or absence of resistant cells.
  • a major objective of the present invention in its broadest aspect is to provide a method that permits that a given sample can be analysed for the presence or absence of microoganisms that are resistant to at least one antimicrobial compound within a relatively short period of time and which is less labour and cost intensive than currently available methods. Being rapid and requiring less labour and costs to be performed, the method is particularly useful for screening of a large number of samples, in particular of food and feed samples.
  • microbial cell is used to designate any type of unicellular organism that can develop resistance to antimicrobial compounds.
  • this term encompasses gram positive and gram negative bacterial cells including cells of bacterial species that are pathogenic or potentially pathogenic for humans and animals.
  • microbial cells include multiresistant strains or clones of such bacterial species.
  • strains of fungal species including filamentous fungi and yeast species, and protozoan species are also encompassed by the invention.
  • resistant indicates that a given strain or clone of the preselected species or group of species is less susceptible to a given antimicrobial compound than the majority ( > 50%) of randomly isolated strains of that or these species, the term “susceptible” indicating that the growth and/or metabolic activity of the microbial strain is inhibited or reduced in the presence of an amount of the antimicrobial at which the majority of randomly isolated strains are not adversely affected in respect of growth and/or metabolic activity.
  • a “resistant” strain as referred to herein includes a strains that is a spontaneous mutant or is mutated deliberately in at least one gene which mutation confers resistance to the given antimicrobial compound to the strain.
  • the term also refers to a strain of a pre-selected species that has acquired resistance to one or more antimicrobial compounds by uptake and/or chromosomal integration of a plasmid or any other episomal elements that carry genes conferring resistance to the antimicrobial compound(s).
  • the method of the invention permits the detection of multiresistant microbial cells, the term "multiresistant” indicating that the cells are resistant to at least two compounds not belonging to the same group of antimicrobial compounds, such as at least three or four of such compounds, e.g. at least five such compounds.
  • the expression "antimicrobial compound” refers to any compound that has a detectable adverse effect on a given type of microbial cells.
  • this term includes the large group of compounds generally referred to as antibiotics which or used in the prevention and therapy of pathogenic microorgansms in both veterinary and human medicine and/or as growth promoters in animal husbandry e.g. in the pig, cattle and poultry industry.
  • antibiotics which or used in the prevention and therapy of pathogenic microorgansms in both veterinary and human medicine and/or as growth promoters in animal husbandry e.g. in the pig, cattle and poultry industry.
  • metal compounds including salts that have antimicrobial activity such as e.g. Hg compounds, and compounds used for disinfection purposes and as preservatives in the food industry.
  • a particularly important objective of the invention is to provide the means of screening large numbers of samples for the presence/absence of multiresistant pathogenic organisms.
  • the requirement for such a mass screening is very evident in the case of carcasses or cut meat or offal products of slaughtered animals.
  • a regulatory demand for mass screening exist for certain types of resistant organisms such as of multiresistant Salmonella.
  • the screening can be performed within a relatively short period of time such as 48 hours or less, preferably within 36 hours or less or even within 24 hours or less.
  • a negative result of the screening procedure according to the invention permits that the carcasses or the cut meat or offal products can be released from the custody. Conversely, if multiresistant microorganisms are detected, the carcasses or the cut meat or offal products can be destroyed or processed in such a manner that the risk of transmitting multiresistant microorganisms to the consumer is eliminated.
  • a selective enrichment step is carried out, this step comprising subjecting the sample to be tested to at least one first enrichment step in a medium and under conditions that will permit selective enrichment of both resistant and non-resistant cells of the pre-selected species or groups of species, followed by subjecting the thus enriched sample to at least one second enrichment step in a medium and under conditions that will permit selective enrichment of resistant cells of the pre-selected species or group of species.
  • the first enrichment step is carried out using a medium and/or conditions that permit(s) cells of this genus to grow while cells belonging to other groups of microorganisms are substantially suppressed in their growth. In this manner, even a small initial number of Salmonella species cells is capable of being amplified selectively at the expense of other cells, i.e. the enrichment medium with the sample to be tested is enriched with respect to Salmonella cells.
  • the medium used in that first enrichment step is typically a liquid, semisolid or solid medium containing sufficient amounts of nutrients for growth and/or metabolic activity of Salmonella cells but supplemented with one or more compounds that permit(s) growth of Salmonella cells of substantially any species or serotype whilst at the same time suppressing most or all cells of other microbial genera.
  • Such selective media for the enrichment of Salmonella are well known to those of skill in the art. Typical examples of such selective media are e.g. found in EP 0 701 624 B1 to which there is specifically referred in this connection.
  • the first enrichment step may also include that the cultivation conditions are selected so as to favour growth of Salmonella at the expense of other microbial cells.
  • a temperature in the range of 39 to 43°C will allow most Salmonella species or serotypes to grow whilst it will suppress many other types of microorganisms thus contributing to enrichment of Salmonella cells.
  • the period of time for the first enrichment step depends on the type of product and the expected level of contamination with the multiresistant organism searched for. Typically, the period of time is in the range of 1 to 20 hours such as e.g. in the range of 8 to 1 5 hours.
  • the above initial selective enrichment step may optionally be preceded by a step where the sample, prior to being selectively enriched, is subjected to a step of non- selective resuscitation of viable but sublethally damaged cells.
  • a resuscitation step that is typically required for samples which are rehydrated, frozen or which contain substances that confer to the sample product a low water activity such as a salt or a sugar is normally carried out in a non-selective buffered medium for a few hours such as from about 2 to about 10 hours, e.g. from 3 to 8 hours.
  • first enrichment step in a first enrichment medium will, when Salmonella cells are present in the sample, lead to a selective enrichment of both resistant and non-resistant cells of Salmonella cells.
  • selective enrichment procedures for Salmonella it is possible, within the time period indicated above, to obtain an enrichment of Salmonella cells that results in the presence of detectable amounts of such cells even if the sample being tested has an content of e.g. 1 cell per g or even less.
  • the thus potentially Salmonella enriched medium is subjected to at least one second enrichment step in a medium and under conditions that will permit selective enrichment of resistant cells of Salmonella.
  • the medium contains one or more antimicrobial compounds in amounts the imply that enriched Salmonella cells that are not resistant to the compound(s) will be prevented from growing or even killed whereas those Salmonella cells that are resistant to the compound(s) will not be inhibited in respect of growth.
  • the second enrichment medium may be the same medium as that used in the first enrichment step but supplemented with the relevant antimicrobial compound(s) at levels where a majority of randomly isolated strains of Salmonella are inhibited in respect of growth.
  • the first and the second enrichment step of the method according to the invention is carried out with the sample present in the same selective medium to which there is added the appropriate amounts of antimicrobial compound(s) at a point in time where the medium is sufficiently enriched in respect of Salmonella. It is also within the scope of the invention that all or a part of the possibly enriched culture resulting from the first enrichment step is transferred to a fresh second enrichment medium containing the relevant antimicrobial compounds.
  • a fresh second enrichment medium containing the relevant antimicrobial compounds.
  • Such an antibiotically spiked fresh medium may or may not contain compounds that renders the medium selective for the species or group of species that are searched for.
  • the period of time for that second enrichment step is typically in the range of 3 to 20 hours such as e.g. in the range of 8 to 1 5 hours. Preferably, the period of time is 1 0 hours or less.
  • the types and the amounts of antimicrobial compounds, such as antibiotics, for the selective medium in the second enrichment step are selected on the basis of the resistance pattern of the particular microorganism(s) the presence/absence of which is to be detected.
  • the enrichment medium may contain all five of these antibiotics.
  • the method permits the presence/absence of organisms that have acquired resistance to preserving agents such as e.g. benzoic acid and sorbic acid, metal compounds such as e.g. Hg and Cu compounds, disinfecting agents and any other types of microbiocidally or microbiostatically active compounds.
  • preserving agents such as e.g. benzoic acid and sorbic acid, metal compounds such as e.g. Hg and Cu compounds, disinfecting agents and any other types of microbiocidally or microbiostatically active compounds.
  • the second enrichment step as described above is followed by at least one further step of enriching the microbial cells of the pre-selected species or group of species.
  • a further enrichment step may be carried out by using an enrichment medium that is non-selective in respect of the pre-determined organism and/or in respect of resistance to antimicrobial compounds, or the medium can be selective with respect to either of these conditions.
  • a step of detecting whether or not resistant cells are present in the second enrichment medium is carried out.
  • this step is carried out using a method that permits a result to be available within a few hours, such as within 2 hours or even within 1 hour.
  • a preferred method in this step is an assay that is based on the identification and/or quantification of a target molecule or a molecular interaction involving a target molecule. Examples of such assays that are well known to the skilled artisan are disclosed in EP 0 701 624 B1 to which there is referred in this regard. Such examples include: assays involving an immunological reaction, such as an immumonassay, e.g.
  • a radio immune assay RIA
  • an enzyme immune assay EIA
  • an enzyme linked immune sorbent assay ELISA
  • a fluorescence antibody technique FA
  • an immonoassay comprising the use of liposomes, micelles or iiposome-like particles in the determination phase
  • an assay that involves immune immobilisation an assay that involves immune agglutination such as latex agglutination
  • the determination step comprises a DNA/DNA hybridisation step, "a RNA/RNA hybridisation assay or a DNA/RNA hybridisation assay, assays wherein the determination step includes a polymerase chain reaction (PCR) or similar in vitro nucleic acid multiplication steps and assays wherein the determination step includes the use of peptide nucleic acid (PNA) or other modified nucleic acid probes.
  • PCR polymerase chain reaction
  • steps involving the use of lectins or similar assays involving receptor binding steps involving the use of bacteriophages, steps that comprise a metabolic conversion of the target organism that results in a change in the electric resistance or impedance of the medium to be tested, steps including a radiometric method and steps involving a chromotographic separation.
  • a presently preferred method of detecting the presence/absence of resistant microorganisms in the medium of the second enrichment step is an immonoassay using antibodies raised against the organism/s) the presence/absence of which is to be detected using an automated or semi-automated instrument such as the EiaFoss instrument developed and marketed by the applicant, Foss Electric, Hiller ⁇ d, Denmark.
  • the medium Prior to subjecting the second enrichment step medium to any of the above assays, the medium may be subjected to a heat treatment or treatments to obtain killing of the organisms to be detected.
  • the method is designed such that it permits the presence or absence of resistant microbial cells to be detected within 48 hours such as within 30 hours or less, preferaby within 24 hours or less such as within 18 hours or less.
  • samples which can be tested by the method include food samples, animal feed samples, samples from the outer environment and samples derived from an animal including a human being.
  • food sample is to be understood in its broadest sense as any liquid, semisolid, solid and dry (rehydrated) material that is edible including raw materials and additives. Accordingly, food samples include e.g. meat and meat products from any edible animal such as cattle, pigs, poultry and fish, milk and milk-based products, eggs and egg-based products, bakery products, confectionaries, vegetables, fruit and beverages including drinking water.
  • Animal feed which can be examined by the present method include concentrate feedstuffs of animal and plant origin, premixes, feed additives and fresh and preserved fodder crop materials including silage.
  • Environmental samples that can be examined for the presence/absence of resistant organisms include samples of surface water and ground water, soil samples and air samples.
  • An interesting application of the present method is the testing of biological samples from animals and human subjects such as samples of blood, sputum, urine and other excreta that are normally tested in clinical laboratories.
  • the method can be used for the detection of any type of microorganism that can acquire resistance to an antimicrobial compound such as bacterial species and fungal species including a yeast species. It will appreciated that the most important pre- determined target organisms are pathogenic or potentially pathogenic strains and isolates of such organisms.
  • Typical examples of such organisms include, but is not limited to, species or group of species that are selected from Enterobacteriaceae species such as Salmonella species and serotypes and Shigella species, Vibrionaceae species, Listeria species, Campylobacter species, Staphylococcus species, Streptococcus species, Pseudomonas species, Hemophilus species, Bordetella species, Pasteurella species and Moraxella species.
  • the method of the invention is adapted specifically for detecting the absence or presence of Salmonella species or serotypes that are resistant to levels of at least four pharmaceutically active antibiotics of different groups to which said species or serotypes are normally susceptible.
  • Such organisms of interest include the specific strain of Salmonella typhimurium that is referred to in the literature as DT104 M .
  • the presence of resistant microbial cells in the second enrichment step medium can be verified by conventional methods of detecting cells that are resistant to antimicrobial compounds. Such methods that have been described above in general terms are well known to the person skilled in the art.
  • the antibiotics ampicillin, chloramphenicol and tetracycline to which the multiresistant strains of Salmonella are known to be resistant were in the range of 2-8 x 10 2 colony forming units (cfu) per ml.
  • the samples were incubated at 37°C for 24 hrs. After incubation, aliquots of the samples were streaked onto Rambach Agar plates. The samples were then boiled for
  • Raw meat samples were divided into 25 g portions and spiked with overnight pure cultures of Salmonella typhimurium DT104 M , overnight pure cultures of non- multitresistant Salmonella strains and a 1 : 1 mixture of multiresistant and non-resistant Salmonella strains.
  • the inoculation level was 200-500 cfu per 25g.
  • the spiked samples were stored at 5°C for 24 hrs prior to analysis.
  • Each of the spiked samples were introduced into 225 ml of Selective Enrichment Broth I (SEBI) and incubated at 41 °C for 6 hrs. Ampicillin, chloramphenicol and tetracycline were mixed and dissolved in sterile deionized water mixed with ethanol. 225 ⁇ l of the antibiotic solution was added to the enrichment culture after the 6 hrs of incubation, and the samples were incubated further at 41 °C for 1 8 hrs.
  • SEBI Selective Enrichment Broth I

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Abstract

L'invention concerne un procédé de détection, dans un échantillon, de la présence ou de l'absence de cellules microbiennes qui sont résistantes à un composé antimicrobien, au moins. Ce procédé comprend une étape d'enrichissement sélectif comportant une première sous-étape consistant à soumettre l'échantillon à une première étape d'enrichissement, dans un milieu et dans des conditions permettant un enrichissement sélectif à la fois des cellules résistantes et de celles non résistantes de l'espèce ou des groupes d'espèces préalablement choisis, puis une seconde sous-étape consistant à soumettre l'échantillon ainsi enrichi à une étape d'enrichissement, dans un milieu et dans des conditions permettant un enrichissement sélectif des cellules résistantes de l'espèce ou du groupe d'espèces préalablement choisi, ainsi qu'une étape de détection de la présence ou de l'absence de cellules résistantes. Ce procédé permet de détecter l'absence/présence d'organismes résistants, tels que par exemple Salmonella en 24 heures environ.
PCT/DK2000/000076 1999-03-03 2000-02-24 Procede de detection de microbes resistant aux agents antimicrobiens Ceased WO2000052196A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU26589/00A AU2658900A (en) 1999-03-03 2000-02-24 Method to detect antimicrobial agent resistant microbes

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DKPA199900294 1999-03-03
DKPA199900294 1999-03-03

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WO2000052196A1 true WO2000052196A1 (fr) 2000-09-08

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0322591A2 (fr) * 1987-12-01 1989-07-05 Abbott Laboratories Essai pour tester la résistance aux antibiotiques
WO1994028163A1 (fr) * 1993-06-02 1994-12-08 Foss Electric A/S Methode de detection des salmonelles

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0322591A2 (fr) * 1987-12-01 1989-07-05 Abbott Laboratories Essai pour tester la résistance aux antibiotiques
WO1994028163A1 (fr) * 1993-06-02 1994-12-08 Foss Electric A/S Methode de detection des salmonelles

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