METHODS FOR DETECTING ESCHERICHIA COLI AND CARRIERS FOR DETECTING E. COLI

• Main Authors: FURUKAWA SATOSHI, NAKAJIMA KAZUTERU, YAMAZAKI TAKAAKI
• Format: Patent
• Language: English; Japanese
• Published: 07.01.2016
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

PROBLEM TO BE SOLVED: To provide a method for detecting Escherichia coli including non-pathogenic strains and enterohemorrhagic E.coli which may cause food poisoning at the same time with high specificity.SOLUTION: Provided is a method for detecting enterohemorrhagic E. coli and general E. coli including enterohemorrhagic E. coli at the same time, where genomic DNA extracted from a sample is used as a template, and multiplex PCR is carried out to amplify a DNA fragment comprising uridine monophosphate kinase gene (pyrH), a DNA fragment comprising vero toxin type I gene (vtx1) and a DNA fragment comprising vero toxin type 2 gene (vtx2) at the same time for detection of the presence or absence of respective amplified products, which enables detection of the presence or absence of enterohemorrhagic E. coli and the presence or absence of general E. coli including hemorrhagic E. coli in the sample. 【課題】 病原性を持たないものを含む大腸菌と、食中毒の原因となる腸管出血性大腸菌の検出を同時にかつ高い特異性で行うことを可能とする。【解決手段】 腸管出血性大腸菌及び大腸菌（腸管出血性大腸菌を含む）を同時に検出する大腸菌の検出方法であって、試料から抽出したゲノムＤＮＡをテンプレートとして、大腸菌のウリジンモノリン酸キナーゼ遺伝子（pyrH）を含むＤＮＡ断片と、ベロ毒素１型遺伝子（vtx1）を含むＤＮＡ断片と、ベロ毒素２型遺伝子（vtx2）を含むＤＮＡ断片とをマルチプレックスＰＣＲにより同時に増幅させ、それぞれの増幅産物の有無を検出することによって、試料中における腸管出血性大腸菌の有無及び大腸菌の有無を同時に検出する。【選択図】 なし