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1. WO2014157611 - METHOD FOR DETECTING WIDE-RANGE OF E. COLI O-SEROGROUPS USING PCR

Publication Number WO/2014/157611
Publication Date 02.10.2014
International Application No. PCT/JP2014/059097
International Filing Date 28.03.2014
IPC
C12N 15/09 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
C12Q 1/68 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
CPC
C12Q 1/689
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
6888for detection or identification of organisms
689for bacteria
C12Q 2600/16
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
2600Oligonucleotides characterized by their use
16Primer sets for multiplex assays
Applicants
  • 国立大学法人宮崎大学 UNIVERSITY OF MIYAZAKI [JP]/[JP]
Inventors
  • 井口 純 IGUCHI Atsushi
Agents
  • 弁護士法人 衞藤法律特許事務所 ETO LAW & PATENT OFFICE CORP.
Priority Data
2013-06846428.03.2013JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) METHOD FOR DETECTING WIDE-RANGE OF E. COLI O-SEROGROUPS USING PCR
(FR) PROCÉDÉ POUR DÉTECTER UNE VASTE GAMME DE SÉROGROUPES O D'E. COLI EN UTILISANT UNE PCR
(JA) PCR法を用いた大腸菌の広域O血清群判定方法
Abstract
(EN)
[Problem] To provide a fast and highly accurate method for classifying enterohemorrhagic E. coli O-serogroups that can replace conventional serological O-serogroup classification. [Solution] A pool template that contains a plurality of primer sets and a single template that contains a solo primer set are used to perform nucleic acid amplification of an E-coli O-antigen sample and identification of E-coli O-antigen, said primer sets containing a forward primer and a reverse primer that enable nucleic acid amplification of base sequences of an O-antigen sugar chain synthetase gene (wzy) and an O-antigen sugar chain transporter protein gene (wzx or wzm/wzt) which are present in a gene region that governs O-antigen synthesis in a E. coli chromosome.
(FR)
L'objet de l'invention est de fournir un procédé rapide et hautement précis pour classer les sérogroupes O d'E.Coli entéro-hémorragiques pouvant remplacer la classification de groupe O sérologique conventionnelle. A cet effet, l'invention propose une matrice commune contenant une pluralité de kits d'amorces et une matrice unique contenant un kit d'amorce individuelle utilisées pour effectuer une amplification d'acide nucléique d'un échantillon d'antigène O d'E-coli et l'identification d'un antigène O d'E-coli, lesdits kits d'amorces contenant une amorce avant et une amorce inverse permettant une amplification de l'acide nucléique des séquences de base d'un gène de synthétase de chaîne de sucre d'antigène O (wzy) et un gène de protéine de transport de chaîne de sucre d'antigène O (wzx ou wzm/wzt) présents dans une région génique qui régit la synthèse d'antigène O dans un chromosome d'E. coli.
(JA)
【課題】現在行われている血清学的なO血清群分類に代わる,迅速で精度の高い腸管出血性大腸菌O血清群の分類手法の提供。 【解決手段】大腸菌染色体上の,O抗原合成を司る遺伝子領域に存在する,O抗原糖鎖合成酵素遺伝子(wzy)とO抗原糖鎖輸送タンパク遺伝子(wzxまたはwzm/wzt),これら塩基配列の核酸増幅を可能とする,フォワードプライマーおよびリバースプライマーを含むプライマーセット。このプライマーセットを,複数含むプールテンプレートならびに単独で含むシングルテンプレートを用いて,大腸菌O抗原試料の核酸増幅を行い,大腸菌O抗原の特定を行う。
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