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WO1992008805A1 - Procede et necessaire de detection de microorganismes - Google Patents

Procede et necessaire de detection de microorganismes Download PDF

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Publication number
WO1992008805A1
WO1992008805A1 PCT/NL1991/000230 NL9100230W WO9208805A1 WO 1992008805 A1 WO1992008805 A1 WO 1992008805A1 NL 9100230 W NL9100230 W NL 9100230W WO 9208805 A1 WO9208805 A1 WO 9208805A1
Authority
WO
WIPO (PCT)
Prior art keywords
microorganisms
demonstrated
pcr
antibodies
bacteria
Prior art date
Application number
PCT/NL1991/000230
Other languages
English (en)
Inventor
Jan Verhoef
Adriaan Camille Fluit
Ruurd Torensma
Original Assignee
U-Gene Research B.V.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from NL9002493A external-priority patent/NL9002493A/nl
Application filed by U-Gene Research B.V. filed Critical U-Gene Research B.V.
Priority to JP4500542A priority Critical patent/JPH06502534A/ja
Priority to EP91920471A priority patent/EP0630413A1/fr
Publication of WO1992008805A1 publication Critical patent/WO1992008805A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6804—Nucleic acid analysis using immunogens
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6865—Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria

Definitions

  • This invention relates to a method and a kit for
  • Salmonella In clinical samples (suchs as urine, feces, saliva, sputum, serum, blood, biopten and the like) or in foods potentially pathogenic species of bacteria may be present in small numbers among large numbers of contaminating other species of microorganisms. These potentially pathogenic bacteria are demonstrated by using selective media.
  • the classical method of demonstrating Salmonella consists in a culture on Salmonella/Shigella agar and enrichment in selenite agar, followed by a biochemical and seriological
  • microorganisms to be determined are first isolated from the sample.
  • a biological medium in which pathogenic microorganisms with a specific antigenic determinant are present is contacted with a magnetic gel to which antibodies directed against this
  • the magnetic particles are then picked up with a magnetic rod and inoculated on an agar plate to form an imprint of about 1 cm in cross-section.
  • the agar plate is incubated so that colonies (as large as the imprint) can be formed.
  • a classical bacteriological method can be used to identify the bacteria in the colony.
  • the immunochemical detection has the further drawbacks that it is hard to distinguish related microorganisms with antisera and that no better sensitivity than 10 6 cells/ml can be realized.
  • a recess is made in the agar at a short distance from the colonies, which recess is filled with a specific antiserum.
  • the colonie are lysed. Diffusion of antigen from the colony and antibody from the recess filled with antiserum leads to an immune precipitate which may then be stained. This procedure takes about two days.
  • HAV hepatitis A virus
  • microorganisms such as Salmonella bacteria.
  • viral infection In viral
  • PCR a number of protocols are described for PCR on pure cultures.
  • the bacteria are pelleted in a centrifuge, included in water and heated for 10 min at 95°C, followed by addition of proteinase K and incubation for 10 min at 55°C.
  • the proteinase is deactivated within 15 min at 95°C, after which RNase is added and the incubation is continued for 15 min at 37°C. Only then is the PCR reaction carried out (Barry et al, Biotechnology 8, 1990, 233-236).
  • the starting material is even DNA which, after lysis and proteinase K treatment, is first purified via phenol extractions and optionally CsCl gradients (Bernet et al, J. Clin. Microbiol. 27, 1989, 2492-2496; Wilson et al, J. Clin. Microbiol. 28, 1990, 1942-1946; Plikaytis et al, J.
  • proteinase K is deactivated by 5 min heating to 80°C.
  • PCR is carried out (see Atlas and Bej, in “PCR protocols; a guide to methods and applications", Innis, Gelfand, Sninsky and White eds, Academic Press, San Diego, 399-406).
  • the invention provides a method and a kit which, unlike all the previously proposed techniques, enable microorganisms such as bacteria, fungi and yeasts to be demonstrated in different kinds of samples in a rapid, specific and sensitive manner.
  • the invention relates in a first aspect to a method for demonstrating microorganisms in a sample wherein the
  • microorganisms to be detected are isolated from the sample by means of antibodies having a specific affinity for these microorganisms and a solid support for these antibodies, and then subjecting the DNA of the microorganisms thus isolated to a PCR in which primers specific for the microorganisms to be demonstrated are used.
  • the invention relates to a kit for demonstrating microorganisms in a sample, comprising
  • antibodies having a specific affinity for the microorganisms to be detected which antibodies are bound to a solid support for these antibodies or are provided with an agent by means of which they can be bound to a solid support for the antibodies likewise belonging to the kit, as well as a set of primers for a PCR that are specific for the microorganisms to be
  • the method according to the invention is particularly suited for demonstrating microorganisms consisting of
  • bacteria, fungi or yeasts of a certain genus, a certain species, or a certain strain and the combination of the use of antibodies having a specific affinity for the
  • microorganisms to be detected so as to isolate these very microorganisms from the sample and the use of a set of primers for a PCR that are specific for the microorganisms to be demonstrated so as to amplify the very DNA of these
  • microorganisms ensures that, e.g., in bacteria the method can distinguish between different genera or between different species, or even between different strains.
  • the invention is not limited to methods for demonstrating pathogenic bacteria in clinical samples (suchs as urine, feces, sputum, saliva, serum, blood, biopten and the like) or foods but also comprises methods that enable identification of, e.g., the yeasts and/oder fungi used for the preparation of consumable products such as bread, beer, wine, cheese, yoghurt and the like.
  • the invention is utilized to demonstrate pathogenic bacteria, which is not possible with the prevailing techniques in a satisfactory manner.
  • the invention is particularly suited for demonstrating microorganisms consisting of bacteria of one of the genera Salmonella, Klebsiella or Listeria. Partly in view of the preference for the use of the invention to determine bacteria the further description of the invention will often refer to bacteria, for the sake of convenience, without contemplating to limit the invention in this respect.
  • microorganisms preferably, monoclonal
  • antibodies are capable of reacting with still living
  • bacteria i.e. have a specific affinity for and can bind to the still living bacteria.
  • still living bacteria that contain DNA
  • the final detection occurs by means of the bacterial DNA.
  • the isolation of the microorganisms to be demonstrated from the sample monoclonal antibodies are used which have been generated against the living microorganism, i.e. that the test animal used as source for the antibodies (mostly a mouse) has been immunized with the living bacteria (at least bacteria closely approaching this condition).
  • the test animal used as source for the antibodies mostly a mouse
  • the living bacteria at least bacteria closely approaching this condition.
  • Salmonella react with dead bacteria only, which means that the sample containing the bacteria must be boiled first. This generally applies to all presently available Immunological assays for the detection of Salmonella. Boiling these
  • adjuvants are used to obtain an immune response. These adjuvants, however, have the drawback that the native proteins of the organism in the adjuvant denature so that many epitopes are destroyed.
  • the bacterial strain is treated with a low concentration formalin.
  • the ingestion of the bacteria by phagocytes pathogenic bacteria are often ingested by phagocytes to live on intracellularly and
  • the preparation of the monoclonal antibodies immunization with the living microorganisms is used in combination or after treatment with an antibiotic (in an amount inhibiting growth of the bacterium) or formaldehyde (in an amount inhibiting ingestion of the bacterium by phagocytes).
  • the antibodies used for the isolation of the microorganisms to be demonstrated must be or become bound to a solid support (optionally after reaction with the microorganisms present in the sample if the
  • antibodies and solid support are suitably modified so as to enable realization of such a later coupling, e.g., via a biotin/avidin system, a hapten/antihapten system etc.).
  • the nature of the solid support is basically not subjected to special restrictions and the solid support may consist of, e.g., inert particles of metal, plastic or glass that can be separated from the rest of the sample by
  • the microorganisms to be demonstrated are isolated from the sample.
  • the microorganisms to be demonstrated are isolated from the sample by adding to the sample magnetic particles to which are bound monoclonal antibodies having specific affinity for the microorganisms to be demonstrated and separating after incubation the magnetic particles with, if present, the microorganisms to be demonstrated bound thereto.
  • magnetic particles with monoclonal antibodies bound thereto having specific affinity for the microorganisms to be demonstrated, which have been obtained by incubating the monoclonal antibodies with magnetic particles coated with antibodies having specific affinity for the monoclonal antibodies. If the monoclonal antibodies are immunoglobulins, e.g., from the mouse, the magnetic particles may be coated with, e.g., immunoglobulins of the goat having specificity for immunoglobulins of the mouse.
  • Bacteria and other microorganisms show the effect of aspecific binding or attachment to solid surfaces. This particularly constitutes a great problem if, in addition to the microorganisms to be demonstrated, the sample to be examined also contains other species of bacteria, such as
  • the invention it is therefore preferred to isolate the DNA from the organisms bound to the magnetic particles and to subject it to PCR in the absence of the magnetic particles.
  • the invention demands little of the sample recovery for PCR; using lysis buffer, phenol extractions and CsCl gradients is not necessary, not even if the method is directed to demonstrating, e.g., Enterobacteriaceae or Listeria.
  • the PCR conditions are known per se to a skilled worker (see, e.g., U.S. patent 4,683,202), as are the species of DNA polymerase suitable therefor, such as the frequently used Taq DNA polymerase.
  • the selection of the primers is specific for the present method: by this selection it can be ensured that the method is specific for a certain genus, a certain species or even a certain strain of a microorganism.
  • suitable primers is not always very easy. This proves to be a problem, e.g., if specificity for a certain genus of bacteria is to be ensured, e.g., for the genus Salmonella, i.e.
  • rRNA primers are a possibility comprised by the invention, but is not suitable in all cases. For particularly in case of
  • Salmonella a rather strong sequence variation proves to occur in the rRNA sequences yet.
  • a suitable alternative has been found in the sequence of the "origin of DNA replication" of the bacterial chromosome (oriC).
  • oriC bacterial chromosome
  • genus Salmonella conserved within, e.g., the genus Salmonella and may therefore serve as the basis for primers for a genus-specific PCR.
  • the invention therefore comprises methods wherein primers are used in PCR that are based on portions of an rRNA
  • the invention also comprises methods wherein primers are used in PCR that are based on portions of a sequence of the origin of DNA replication.
  • a concrete example thereof is a method wherein bacteria of the genus Salmonella are demonstrated and the following oligonucleotide primers are used in PCR:
  • primer 1 5'-TTATTAGGATCGCGCCAGGC-3'
  • primer 2 5'-AAAGAATAACCGTTGTTCAC-3'.
  • a second example is a method wherein bacteria of the genus Klebsiella are demonstrated and the following
  • oligonucleotide primers are used in PCR:
  • primer 1 5'-CTTGTCTTGTGGATAAGTCA-3 ,
  • primer 2 5'-TCTTCTGTGGATAACTATGC-3' It has been established by way of experiment that with this primer combination and with Klebsiella-specific
  • the method according to the invention is selective for bacteria of the genus Klebsiella, which means a positive result for Klebsiella oxvtoca and Klebsiella
  • Citrobacter freundii Serratia marcescens, Proteus mirabilis, Enterobacter cloacae and Enterobacter aerogenes.
  • primer 1 5'-CTTGTCTTGTG ⁇ GATAAGTCA-3'
  • primer 2 5'-AAGTATAACCGTTGCCTGA-3'
  • MAB 55-23-B7 ECACC 90112807
  • MAB 70-2-2A ECACC 90112808
  • MAB 71-40-A1 ECACC 90112809
  • the PCR technique was used for the amplification of a 160 bp portion of the sequence of the origin of DNA replication.
  • primers were used having a sequence derived from the origin of replication of Salmonella typhimurium.
  • oligonucleotide primers were synthesized on a Pharmacia LKB Gene Assembler Plus Synthesizer using the phosphoramidite technology. After deprotection and cleavage from the solid support the oligonucleotides were purified by chromatography on NAP10 columns.
  • the primers had the following sequences: primer 1: 5'-TTATTAGGATCGCGCCAGGC-3'
  • primer 2 5'-AAAGAATAACCGTTGTTCAC-3'
  • the specificity of the set of primers was tested in a PCR at 27 Salmonella strains and 19 other Enterobacteriaceae (no salmonella) (see Table A).
  • the PCR reaction mixture (final volume 100 ⁇ l) consisted of 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 1.5 mM MgCl 2 , 0.001% (w/v) gelatin, 100 ⁇ M of each dNTP, 0.5 ⁇ M of each primer, 1.25 U Ampli-Taq polymerase (Cetus, Emeryville, CA, USA).
  • the amplification was carried out in 25 cycles on a Perkin-Elmer Thermocycle (Perkin-Elmer
  • Enterobacteriaceae such as Shigella, Escherichia coli,
  • Citrobacter and Psendomonas it was derived therefrom that the selected set of primers is Salmonella-specific.
  • immunoglobulins of the goat which were specific for igG and IgM of the mouse.
  • the binding of monoclonal antibodies (of subclass IgM), directed against Salmonella bacteria, to the magnetic beads occurred by incubating supernatant of the hybridoma culture with the coated magnetic beads for 5 min at 35°C in a phosphate buffered saline with 1% gelatin (gPBS).
  • the magnetic particles were isolated by means of a magnet, after which the supernatant was thrown away.
  • test samples were suspended in gPBS and added to the magnetic particles. After incubation of 15 min at 35°C the magnetic particles were isolated by means of a magnet and washed three times with gPBS. After the last washing step the magnetic particles were resuspended in aqua bidest.
  • the supernatant obtained after incubation with the bacteria was also subjected to PCR. No amplification of the target DNA sequence was found, from which it can be derived that the binding of the Salmonella bacteria to the monoclonal

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Abstract

Procédé de détection de microorganismes consistant dans un premier temps à les isoler de l'échantillon à l'aide d'anticorps et d'un support solide pour ceux-ci, puis, éventuellement après avoir isolé l'ADN des microorganismes liés, à effectuer une réaction en chaîne de polymérase. Ces anticorps sont spécifiques des microorganismes à détecter et de préférence dressés contre des microorganismes toujours vivants. Des particules magnétiques sont tout spécialement aptes à servir de support solide. Dans la réaction en chaîne de polymérase, on utilise des amorces spécifiques des microorganismes à détecter et de préférence basées sur une séquence d'ARNr ou une séquence d'ADN d'origine de réplication.
PCT/NL1991/000230 1990-11-15 1991-11-15 Procede et necessaire de detection de microorganismes WO1992008805A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP4500542A JPH06502534A (ja) 1990-11-15 1991-11-15 微生物を分析する方法及びキット
EP91920471A EP0630413A1 (fr) 1990-11-15 1991-11-15 Procede et necessaire de detection de microorganismes

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
NL9002493A NL9002493A (nl) 1990-11-15 1990-11-15 Werkwijze en kit voor het aantonen van microoerganismen.
NL9002493 1990-11-15
NL9002696 1990-12-07
NL9002696A NL9002696A (nl) 1990-11-15 1990-12-07 Werkwijze en kit voor het aantonen van microoerganismen.

Publications (1)

Publication Number Publication Date
WO1992008805A1 true WO1992008805A1 (fr) 1992-05-29

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PCT/NL1991/000230 WO1992008805A1 (fr) 1990-11-15 1991-11-15 Procede et necessaire de detection de microorganismes

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EP (1) EP0630413A1 (fr)
JP (1) JPH06502534A (fr)
CA (1) CA2095843A1 (fr)
NL (1) NL9002696A (fr)
WO (1) WO1992008805A1 (fr)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995000664A1 (fr) * 1993-06-17 1995-01-05 Bioteknologisk Institut Identification de salmonella par amplification enzymatique
EP0605003A3 (fr) * 1992-12-31 1995-06-07 Vicam Lp Méthode d'essai pour détecter la présence de bactéries.
WO1999029892A1 (fr) * 1997-12-05 1999-06-17 The Perkin-Elmer Corporation Procede de detection d'organismes dans un echantillon
WO2003012447A3 (fr) * 2001-07-23 2003-08-07 Inst Chemo Biosensorik Dispositif et procede pour la mise en evidence de micro-organismes, de cellules eucaryotes ou d'organites vivants
EP1767651A1 (fr) * 2005-09-26 2007-03-28 Ludwig-Maximilians-Universität München Séquences d'acide nucléique spécifiques d'oriC et leur utilisation
WO2008116941A1 (fr) * 2007-03-26 2008-10-02 Fundación Gaiker Méthode et dispositif pour la détection d'un matériel génétique au moyen d'une réaction en chaîne par polymérase
US8710836B2 (en) 2008-12-10 2014-04-29 Nanomr, Inc. NMR, instrumentation, and flow meter/controller continuously detecting MR signals, from continuously flowing sample material
US8841104B2 (en) 2010-04-21 2014-09-23 Nanomr, Inc. Methods for isolating a target analyte from a heterogeneous sample
US9389225B2 (en) 2010-04-21 2016-07-12 Dna Electronics, Inc. Separating target analytes using alternating magnetic fields
US9428547B2 (en) 2010-04-21 2016-08-30 Dna Electronics, Inc. Compositions for isolating a target analyte from a heterogeneous sample
US9476812B2 (en) 2010-04-21 2016-10-25 Dna Electronics, Inc. Methods for isolating a target analyte from a heterogeneous sample
US9551704B2 (en) 2012-12-19 2017-01-24 Dna Electronics, Inc. Target detection
US9599610B2 (en) 2012-12-19 2017-03-21 Dnae Group Holdings Limited Target capture system
US9804069B2 (en) 2012-12-19 2017-10-31 Dnae Group Holdings Limited Methods for degrading nucleic acid
US9902949B2 (en) 2012-12-19 2018-02-27 Dnae Group Holdings Limited Methods for universal target capture
US9995742B2 (en) 2012-12-19 2018-06-12 Dnae Group Holdings Limited Sample entry
US10000557B2 (en) 2012-12-19 2018-06-19 Dnae Group Holdings Limited Methods for raising antibodies
CN113567665A (zh) * 2021-08-16 2021-10-29 固安林科特生物工程有限公司 一种用于沙眼衣原体抗原检测的裂解液及检测方法

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FR2537725A1 (fr) * 1982-12-09 1984-06-15 Pasteur Institut Procede de detection immunobacteriologique de germes pathogenes dans des milieux biologiques contamines
EP0145356A2 (fr) * 1983-11-22 1985-06-19 National Research Development Corporation Méthode d'essai d'ADN ou ARN à partir de particules virales
EP0366448A2 (fr) * 1988-10-25 1990-05-02 The General Hospital Corporation Procédé pour détecter et identifier des moitiés contenant des acides nucléiques
EP0395292A2 (fr) * 1989-04-20 1990-10-31 Thomas Gerard Barry Génération de sondes spécifiques pour les séquences nucléotidiques cibles
EP0402997A2 (fr) * 1989-06-15 1990-12-19 Akzo Nobel N.V. Procédé pour déterminer des acides nucléiques
WO1991008308A1 (fr) * 1989-11-30 1991-06-13 Pharmacia Genetic Engineering, Inc. Nouvelle methode permettant de detecter une sequence d'acide nucleique specifique a partir d'un prelevement de cellules

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FR2537725A1 (fr) * 1982-12-09 1984-06-15 Pasteur Institut Procede de detection immunobacteriologique de germes pathogenes dans des milieux biologiques contamines
EP0145356A2 (fr) * 1983-11-22 1985-06-19 National Research Development Corporation Méthode d'essai d'ADN ou ARN à partir de particules virales
EP0366448A2 (fr) * 1988-10-25 1990-05-02 The General Hospital Corporation Procédé pour détecter et identifier des moitiés contenant des acides nucléiques
EP0395292A2 (fr) * 1989-04-20 1990-10-31 Thomas Gerard Barry Génération de sondes spécifiques pour les séquences nucléotidiques cibles
EP0402997A2 (fr) * 1989-06-15 1990-12-19 Akzo Nobel N.V. Procédé pour déterminer des acides nucléiques
WO1991008308A1 (fr) * 1989-11-30 1991-06-13 Pharmacia Genetic Engineering, Inc. Nouvelle methode permettant de detecter une sequence d'acide nucleique specifique a partir d'un prelevement de cellules

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Proc. Natl. Acad. Sci., volume 87, April 1990 (Washington, US) R.W. Jansen et al.: "Molecular epidemiology of human hepatitis A virus defined by an antigen-capture polymerase chain reaction method", pages 2867-2871, see the whole article (cited in the application) *

Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5491068A (en) * 1991-02-14 1996-02-13 Vicam, L.P. Assay method for detecting the presence of bacteria
US5695946A (en) * 1991-02-14 1997-12-09 Vicam, Lp Assay method for detecting presence of bacteria
EP0605003A3 (fr) * 1992-12-31 1995-06-07 Vicam Lp Méthode d'essai pour détecter la présence de bactéries.
WO1995000664A1 (fr) * 1993-06-17 1995-01-05 Bioteknologisk Institut Identification de salmonella par amplification enzymatique
US6004747A (en) * 1993-06-17 1999-12-21 John Elmerdahl Olsen Salmonella identification by the polymerase chain reaction
WO1999029892A1 (fr) * 1997-12-05 1999-06-17 The Perkin-Elmer Corporation Procede de detection d'organismes dans un echantillon
WO2003012447A3 (fr) * 2001-07-23 2003-08-07 Inst Chemo Biosensorik Dispositif et procede pour la mise en evidence de micro-organismes, de cellules eucaryotes ou d'organites vivants
EP1767651A1 (fr) * 2005-09-26 2007-03-28 Ludwig-Maximilians-Universität München Séquences d'acide nucléique spécifiques d'oriC et leur utilisation
WO2007039182A1 (fr) * 2005-09-26 2007-04-12 Ludwig-Maximilians-Universität München SÉQUENCES D'ACIDES NUCLÉIQUES SPÉCIFIQUES À oriC ET UTILISATION DE CELLES-CI
WO2008116941A1 (fr) * 2007-03-26 2008-10-02 Fundación Gaiker Méthode et dispositif pour la détection d'un matériel génétique au moyen d'une réaction en chaîne par polymérase
US8710836B2 (en) 2008-12-10 2014-04-29 Nanomr, Inc. NMR, instrumentation, and flow meter/controller continuously detecting MR signals, from continuously flowing sample material
US11448646B2 (en) 2010-04-21 2022-09-20 Dnae Group Holdings Limited Isolating a target analyte from a body fluid
US9389225B2 (en) 2010-04-21 2016-07-12 Dna Electronics, Inc. Separating target analytes using alternating magnetic fields
US9428547B2 (en) 2010-04-21 2016-08-30 Dna Electronics, Inc. Compositions for isolating a target analyte from a heterogeneous sample
US9476812B2 (en) 2010-04-21 2016-10-25 Dna Electronics, Inc. Methods for isolating a target analyte from a heterogeneous sample
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CA2095843A1 (fr) 1992-05-16

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