WO1996017961A1 - Procede et dispositif de detection de bacteries - Google Patents

Procede et dispositif de detection de bacteries Download PDF

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Publication number
WO1996017961A1
WO1996017961A1 PCT/JP1995/002475 JP9502475W WO9617961A1 WO 1996017961 A1 WO1996017961 A1 WO 1996017961A1 JP 9502475 W JP9502475 W JP 9502475W WO 9617961 A1 WO9617961 A1 WO 9617961A1
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WIPO (PCT)
Prior art keywords
phage
detected
bacterium
bacteria
detection method
Prior art date
Application number
PCT/JP1995/002475
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English (en)
Japanese (ja)
Inventor
Sinji Tuyumu
Original Assignee
Tohoku Electronic Industrial Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tohoku Electronic Industrial Co., Ltd. filed Critical Tohoku Electronic Industrial Co., Ltd.
Publication of WO1996017961A1 publication Critical patent/WO1996017961A1/fr

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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
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • 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
    • 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 emission m is applied to a method for detecting bacteria and a method for detecting this]]].
  • c file temporary is ji Ri virus der to be infected with fine m, municipal district also DN ⁇ is It takes a form in which a nucleic acid composed of RNA is coated with a structure composed of protein.
  • the phage value is usually the total number of the phage particles in the sample. In this method, an increase in the phageka value indicates the presence of the target bacteria.
  • Reporter genes are exogenous genes that are not present in the organism being measured.
  • ⁇ ⁇ ⁇ is a marker-gene.
  • the selectable marker gene used for the selection of the transformant is different from the selected marker gene, and refers to a gene that can be used for the purpose of quantitatively or histologically measuring the specific gene.
  • the reporter ⁇ iz ⁇ is analyzed by measuring the activity of the reporter ⁇ iz ⁇ into a dye or brassamide.
  • the phage method is formed by mixing a sample with a previously cultivated susceptible cell i and cultivating it for 11 n times in order to measure the phage titer of the sample after phage addition.
  • the number of plaques must be measured. For this reason, it takes time [ ⁇ ] before spinning.
  • complicated operations are required for measurement, and the measurement must be refined.
  • the infection may not be established. I can't do it.
  • the EL 1SA method it is necessary to wash the bacteria several times after fixing the bacteria, and at that time, the bacteria are easily washed away.
  • the method of transferring the revoter gene is as described above, and the revoter-transducer is incorporated into a bacterial stain (or brassmid) and its activity is measured. Although it is suitable for monitoring bacteria that have incorporated a reporter gene, it cannot be used to detect the same bacteria in nature. If incorporated, the number of copies is small and the reporter
  • ⁇ i 3 ⁇ 4 ⁇ is not enough.
  • the ⁇ may refer to the ⁇ ⁇ and ⁇ ⁇ genes, and at the end of this process, the ⁇ ' ⁇ [will appear. There is.
  • each of the conventional j-power-methods has excellent characteristics. There are many suitable ones. However, the diagnosis of qi caused by bacteria, the examination of fine apricots as part of a survey, the examination of foods, and the examination of fines from humans during a period of time, etc., were carried out on a long-term basis. There are only a limited number of methods that can be used in the method. Vi3 ⁇ 4) Both are inadequate. For this reason, a smaller number of There is a need for a detection method that has high sensitivity that can detect even a specific object, and that has high accuracy that can specifically detect a specific i. Denote i! Ij m
  • An object of the present invention is to provide a method for detecting bacteria that is capable of processing a large amount of sample in a short period of time and has excellent sensitivity and accuracy.
  • this detection method can use the above detection method to process large quantities of samples in a short period of time, and also to develop a bacterial detection device with excellent sensitivity and accuracy. Be mouthful.
  • a labeled phage mainly containing the bacterium to be detected is added to a subject, and the signal generated on the label is measured 1. This is the feature.
  • phage is a virus that infects cells, but its host paradigm is five and can only be sensed in particular ways. Therefore, the phage that flows in the detection method using 3 ⁇ 4 ⁇ ) is ⁇ - finely sought to be detected, and is selected according to the bacteria.
  • large intestine contains lambda phage, ⁇ phage, and ⁇ 13 phage, and CV 1 and CP 2 against Xanthomonas camp estx is v.
  • Citri Citri
  • ⁇ 105 Citri
  • i with D 3 VF-13 can be used for Vibrio fetus.
  • the phage can be labeled by direct labeling of the phage itself with the radioisotope (RI) element labeling method, or by the transfer of a reporter gene to the phage's own nucleic acid (hereinafter sometimes referred to as phage nucleic acid). This can be done by incorporating it. Labeling by direct labeling can be measured directly.However, when a reporter gene is incorporated, the reporter gene is expressed and the production of the encoded protein is detected. Can be measured indirectly.
  • RI radioisotope
  • a phage coat protein can be formed using a radioactive isotope-containing amino acid to incorporate a radioisotope-containing nucleotide into a phage nucleic acid. I just need.
  • phage is labeled with phage-biotin, and then ⁇ -labeled avidin (or streptavidin) is added to avidin (where ⁇ stands for streptavidin). It forms a solid ⁇ , which solidifies ⁇ on the phage.
  • avidin or streptavidin
  • antigen-antibody reactions can be performed.
  • the element that can be used here can be any element that can be used in this branch town, such as alkaline phosphatase and herb oxidase. It can be.
  • one of the revoter genes that accumulates in the phage nucleus is: For example, the opin synthesis ⁇ gene, the chloramphenicol acetyltransferase (CAT) gene, the NTP II gene, the gene for glucuronidase (GUS), the ⁇ -galactosidase (L ac Ii) Genes and luciferase genes such as firefly luciferase (Luc) and the marine luminescent bacterium Vibrio luciferase (LuX). These revoter genes are integrated into the phage nucleic acid such that they are transposon-bound and cloning of the phage are controlled by the phage mouth motor.
  • CAT chloramphenicol acetyltransferase
  • NTP II the gene for glucuronidase
  • GUS glucuronidase
  • L ac Ii ⁇ -galactosidase
  • luciferase genes
  • the phage thus labeled is added to the subject II to infect bacteria contained in the width of the subject. If there is a cell i to be detected in the test subject, the phage adsorbs to the bacterium, and the nucleic acid from the phage is transferred to the cell. Nucleic acids injected into bacteria are replicated by the elements produced by these genes, and spread in a short time. Thereafter, each replicated nucleic acid produces a 1: 1 coat of the phage itself, and the protein coats each phage nucleic acid to form multiple phage particles. These high-speed processes can be performed in a conventional manner.
  • the phage of test ⁇ 4I 'and the phage remaining without fine adsorption are separated by a suitable T-stage such as a filter, and labeled from the separated phages.
  • a suitable T-stage such as a filter
  • the detection of ⁇ coming in the mountains is performed.
  • the test i-blu contains a cell as I I ii'j ⁇
  • the signal coming from the separated cell j i to the phage labeling is detected.
  • the ⁇ sign to be detected and the detection method depend on the type of sign Different.
  • the specific radiation emitted by the radioisotope may be measured by a scintillation counter or the like.
  • the substrate of the labeled enzyme may be added, and specific color development, luminescence, etc. may be measured with a colorimeter or a photometer.
  • the measurement is usually performed at the stage when the phage is adsorbed on the bacteria.
  • the phage nucleic acid incorporates the reporter gene
  • the phage is infected with bacteria, and the phage nucleic acid containing the reporter gene is actively replicated, transcribed and translated, and then the activity of the reporter gene is measured.
  • the time required from the infection of the phage to the measurement may be as long as the fuzz is wide enough to be detectable, and is not particularly limited.
  • the nature of the reporter gene is determined by the fact that the reporter gene is expressed and that the gene encoded by the gene is produced. Production: If the ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ to be produced is ⁇ primary as described above, it is necessary to detect each ⁇ ;
  • the phage which is a virus that infects cell j, has a very small ⁇ zi £ i H, and can only infect cell ji in particular r. In other words, it has special characteristics for details. For this reason, if there is a fine j ⁇ ⁇ ⁇ i in the width of the subject that contains the phage, the phage is infected with the fine l3 ⁇ 4i, and if there is no fine ⁇ ⁇ ⁇ ' ⁇ l, il! I It floats in the subject bub without being infected by any bacteria. Therefore, by targeting phage to Icheon, the target can be improved. Detection with high accuracy is possible.
  • a phage When a phage encounters a host bacterium, it first adsorbs to the bacterium and injects the phage's nucleic acid into cells. Nucleic acids injected into bacteria are replicated by enzymes produced by these genes and amplified several hundred times in a short time. Then, each replicated nucleic acid is phage! ⁇ Produces a coat protein of the body: S, which coats each phage nucleic acid to form a large number of phage particles. Therefore, even if the number of cells to be detected is small, the measurement is easy because the measurement is performed on phages that are several to several times wider, and the detection sensitivity can be greatly increased. .
  • the method for detecting fine cocoons according to this method is capable of processing a large amount of sample in a short time [5/5], and is a convenient method with excellent sensitivity and accuracy.
  • the operation is not required to be performed in a shorter time (? ⁇ ] (A ⁇
  • the phage method does not allow the phage to be measured without the phage J even if the phage is sucked into the cell and the phage nucleic acid is damaged.
  • this method using 3 ⁇ 4m it is possible to detect even if the phage is only absorbed finely by labeling the phage
  • the ir-converted fine particles flow into the wash ⁇ , and] ' ⁇ ⁇ ⁇ :: in] m is generated.
  • /] does not need to standardize the details ii, and does not reject such ⁇ ⁇ m.
  • the preparation of labeled phage which can be performed by the method according to Ili1, is easy, and the preparation of the labeled phage can be performed by the EL1S method. It does not require a great deal of effort, such as the preparation of antibodies to be performed.
  • the conventional method of incorporating a reporter gene cannot detect only bacteria and their progeny incorporating the reporter gene.
  • the levator gene can be incorporated only into the specific bacteria present in the sample block. For this reason, it is possible to detect a specific fine sprout in a sample with high accuracy, such as detecting a mixed fine cocoon.
  • the fine cocoon detection device comprises a subject processing unit 1 for infecting phage labeled on a fine j3 ⁇ 4i of a subject bub and a label for two phages. It is characterized in that a detector 2 for detecting a signal coming from the mountain and a data processing unit 3 for processing the signal from the detector 2 for this detection I are provided ft.
  • a process m is performed to allow the labeled phage to be sensed in the cell under test.
  • the phage and infection treatment are as described in detail in the above “m” detection method.
  • the labeled phage may be added in advance before the specimen is contained in the specimen processing section 1, or the specimen may be added to the specimen processing section.
  • 1 may be provided with an appropriate phage liquid stage.
  • Detector 2 is for detecting a signal coming to the sign, and is selected according to the rareness of i: for example, A chemiluminescence measuring device using a photoelectric conversion element such as a photomultiplier tube, an R 1 detector, etc. can be used.
  • the detector 2 may be provided integrally with the subject processing unit 1 or may be provided independently of the subject processing unit 1 and move to a predetermined position in the subject processing unit 1 during measurement.
  • the format may be Further, the subject processing unit 1 and the detector 2 may form a line, and the subject processed in the subject processing unit 1 may be transported into the detector 2.
  • Ir measurement is sent to the data processing unit 3 as a normal electrical signal? 0: Will be welcomed.
  • the data processed in the data processing section 3 is output to a display device, a recording device, or the like.
  • the device for detecting fine ji according to the second aspect is suitable for performing the above-described method for detecting fine 3 ⁇ 4i3 ⁇ 4i by using the above-described method, and is capable of processing a large number of samples with short jtil [ ⁇ ]. This makes it possible to perform highly sensitive and accurate measurements. In addition, it is also possible to make the operation of -a ill, so that the measurement ii 3 ⁇ 4'i-can be performed just by moving the subject. ? ⁇ ' ⁇ description of m ⁇
  • Figure 1 outlines the details of the device for detecting a detailed i3 ⁇ 4i using this method.
  • Fig. 2 shows an example of a device for detecting fine i3 ⁇ 4 by means of ⁇ m, which is a chemical spectrophotometer measuring ⁇ ⁇ ; is there. Best form to implement departure
  • phages were purified by ultracentrifugation density gradient method and then labeled with photobiotin. Then, the labeled phage solution was added to a known concentration of E. coli collected on Millipore Filter (Millipore) so as to have a titer of 100 to 1000 times, and the phage was added. Adsorbed. 10 minutes after the addition of the phage solution, the filter is washed with a phosphate buffer or the like, alkali phosphatase-labeled avidin is added, and the phage is adsorbed, followed by addition of fluorescent m (AMPD). The luminescence of the luminescence was measured with a chemical spectrophotometer IT machine. As a result, it was possible to detect human intestine i3 ⁇ 4i of 2 ⁇ 10 3 flA
  • AMPD fluorescent m
  • the same filter was subjected to the same treatment without using Escherichia coli, and the luminescence was measured with a chemiluminescence analyzer in the same manner as a control.
  • i3 ⁇ 4i Xanthomonas campestris v. Citri
  • the disease C1 and c ⁇ 2 which are the phages of the specific Performed in
  • a fine i3 ⁇ 4i of 1 ⁇ 10 : ' ⁇ could be detected.
  • the equipment shown in the ⁇ 2 screen uses a substance that emits chemical light as a phage label in the detection method described above. It can be ffl suitable for the river.
  • This apparatus has a container bottom member 12 having a circular bottom plate provided with a plate on the periphery thereof, a cylindrical container frame member 13 and a storage container 11 composed of a cap. Threads are provided on the bottom wall of the container bottom member 12 and the outer wall near one end of the container frame member 13 so that they can be screwed together.
  • a sealing member 14 made of rubber, plastic, or the like is provided at the lower end of the container frame member 13 to ensure liquid tightness in the integrally formed container 11.
  • the container bottom W12 and the container frame member 13 are made of a normal metal, but may be made of a material having elasticity such as plastic, and in that case, the sealing member 14 may not be provided.
  • the bottom plate of the container bottom member 12 is provided with a valve 15 for freely releasing the liquid in the container 11 and draining the liquid.
  • the vessel bottom member 12 is provided with a heating T-stage (not shown) for heating the container 11, and the temperature is controlled by a temperature control device it 33.
  • a projection 17 for locking the finoletor 16 is provided on the inner wall of the casing member 13, and the filter 16 locked on the projection 17 is provided from above. It is determined by the W fixing member 18;
  • Two wooden liquid guides 23 and 24 are located above part 11 of the container 11. These guides 23 and 24 are connected to the test storage tank 20 and the labeled phage solution 24, respectively, and the test and labeled phage solutions are respectively stored in the container 11 inside. Lead.
  • only two 11 ⁇ M tanks and the introduced Tf connected to them are shown, but if necessary, for example, washing water, etc. Storage tanks can be provided.
  • a photomultiplier tube 25 is provided above the liquid introduction tubes 23 and 24.
  • a light filter 26 and a photomultiplier tube 25 are provided in the case of the photomultiplier tube 25 and the container 11, a light filter 26 and a photomultiplier tube 25 are provided.
  • An optical shutter 27 is provided to protect the light.
  • the photomultiplier tube 25 is covered with a cooling member 28 for reducing noise, and is further connected to a main controller 31 via a preamble 29 and an amplitude discriminator 30.
  • an optical shutter controller 32 for controlling the optical shutter 27 and a heating means for the container 11 are connected to the controller 31.
  • Data processing is performed.
  • the data processed SJ1 in the main control device 31 is output to the display device 34 and the printer 35 for display and recording.
  • the detection of the cell [i] flowing through this device iTc is performed as follows. First, in the ⁇ ) ⁇ step of the measurement, the subject is poured into the test ⁇ storage 3 ⁇ 4 ⁇ 20, and the labeled phage solution is poured into the labeled phage solution W 21.
  • the labeling of the phage which is performed in this manner, is based on chemical light or flash light.
  • the phage labeled with photobiotin based on the ⁇ element labeling method will be described as an example.
  • the container 11 is formed in accordance with U12, and is mounted at a predetermined position in the apparatus.
  • the valve 15 provided on the bottom of the container 11 is released, and then the container ⁇ from the storage tank 20 to be inspected through the guide '? ⁇ ' 23 into the container 11 Injects ⁇ into the subject.
  • the injected test ⁇ The solid containing bacteria and the soluble component are separated by the filter 16, and the filtrate containing the soluble component passes through the filter 16 and is discharged from the valve 15 to the outside of the container.
  • the labeled phage solution is injected into the container 11 from the labeled phage solution storage tank 21 via the introduction tube 24, and brought into contact with the bacteria remaining on the filter. As a result, when the target bacteria are present on the filter, the bacteria are infected with the phage.
  • washing water is introduced into the container by washing water from an introduction system (not shown) to wash the filter, and similarly, a solution of avidin-labeled avidin is introduced from an introduction system (not shown). Fix phosphatase to bacteria. After further washing to remove unreacted avidin, ⁇ ⁇ MPPD, a fluorescent dye, is introduced from a guide system (not shown).
  • the optical element is multiplied by, for example, a radiation measuring instrument.
  • a radiation measuring instrument it is possible to use the ill river in the method of detecting fine teeth described above using 11 as a marker. If this is the case, separate the cells on the filter 16 of the storage container 11; after t A, close the valve 15 and store the scintillation catheter. Inject into the instrument and measure.
  • the measurement period Prflb and the container 11 are fixed directly below the photomultiplier tube 25, but the container 11 is mounted on a moving stage, and until the addition of AMPPD, for example. Can be performed immediately below each storage tank, and at the time of measurement, it can be moved to immediately below the photomultiplier tube 25 to perform measurement. ⁇ Business profit / H possibility
  • the method of detecting fine cocoons by using this method can be widely applied to the detection and identification of specific fine cocoons in various fields such as food, medicine, and industry.For example, sewage, seawater, industrial wastewater, etc. Of cocoons in foods such as assessment of bacteria, juice, beer, etc., prompt diagnosis of infection with specific pathogens in humans and animals, and identification from the perspective of product quarantine ⁇ Suitable for early detection of pathogenic bacteria, such as U1.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Virology (AREA)
  • Toxicology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un procédé de détection de bactéries, avec lequel un phage marqué, qui utilise la bactérie à détecter comme hôte, est ajouté à un spécimen, et les signaux provenant de ce marquage sont mesurés. Un dispositif servant à appliquer le procédé de détection comprend un élément (1) qui infecte le phage marqué avec la bactérie à l'intérieur du spécimen, un détecteur (2) pour détecter les signaux provenant du marquage de ce phage et un processeur de données (3) pour traiter les signaux provenant du détecteur (2).
PCT/JP1995/002475 1994-12-05 1995-12-05 Procede et dispositif de detection de bacteries WO1996017961A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP6/300866 1994-12-05
JP30086694A JP3615810B2 (ja) 1994-12-05 1994-12-05 細菌の検出方法及び検出装置

Publications (1)

Publication Number Publication Date
WO1996017961A1 true WO1996017961A1 (fr) 1996-06-13

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PCT/JP1995/002475 WO1996017961A1 (fr) 1994-12-05 1995-12-05 Procede et dispositif de detection de bacteries

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JP (1) JP3615810B2 (fr)
WO (1) WO1996017961A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE38863E1 (en) 1995-02-03 2005-11-01 Ruy Tchao Chemotaxis assay procedure

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3270722B2 (ja) * 1996-09-27 2002-04-02 オルガノ株式会社 細菌の検出方法及び検出装置
JP2016032435A (ja) * 2014-07-30 2016-03-10 国立大学法人広島大学 バクテリオファージ、カンキツかいよう病菌の検出剤、カンキツかいよう病菌の検出方法、カンキツかいよう病の防除剤およびカンキツかいよう病の防除方法
ES2694874T3 (es) * 2015-11-20 2018-12-27 Sinamira AG Procedimiento y dispositivo para la detección de bacterias

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6143998A (ja) * 1984-06-05 1986-03-03 アマーシャム　インターナショナル　パブリック　リミティド　カンパニー 環境中微生物の検出方法
JPS61272656A (ja) * 1985-05-24 1986-12-02 エンゾ−　バイオケム　インコ−ポレイテツド 分析物成分の検出方法および検出用組成物
WO1988004326A1 (fr) * 1986-12-01 1988-06-16 Mcdonnell Douglas Corporation Procede d'identification des organismes inconnus
JPH0228772A (ja) * 1988-07-18 1990-01-30 Hitachi Electron Eng Co Ltd 微生物の識別方法および識別装置
JPH04293497A (ja) * 1991-03-25 1992-10-19 Hitachi Electron Eng Co Ltd 生菌数測定・菌種同定システム

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6143998A (ja) * 1984-06-05 1986-03-03 アマーシャム　インターナショナル　パブリック　リミティド　カンパニー 環境中微生物の検出方法
JPS61272656A (ja) * 1985-05-24 1986-12-02 エンゾ−　バイオケム　インコ−ポレイテツド 分析物成分の検出方法および検出用組成物
WO1988004326A1 (fr) * 1986-12-01 1988-06-16 Mcdonnell Douglas Corporation Procede d'identification des organismes inconnus
JPH0228772A (ja) * 1988-07-18 1990-01-30 Hitachi Electron Eng Co Ltd 微生物の識別方法および識別装置
JPH04293497A (ja) * 1991-03-25 1992-10-19 Hitachi Electron Eng Co Ltd 生菌数測定・菌種同定システム

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE38863E1 (en) 1995-02-03 2005-11-01 Ruy Tchao Chemotaxis assay procedure
USRE40747E1 (en) 1995-02-03 2009-06-16 Ruy Tchao Chemotaxis assay procedure

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JPH08154700A (ja) 1996-06-18

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