Top > Search of International Patents > BROAD-SPECTRUM METHOD FOR DETERMINING E. COLI FLAGELLAR ANTIGEN TYPE USING PCR METHOD

BROAD-SPECTRUM METHOD FOR DETERMINING E. COLI FLAGELLAR ANTIGEN TYPE USING PCR METHOD

Foreign code F180009494
File No. S2016-0931-C0
Posted date Nov 1, 2018
Country WIPO
International application number 2017JP025817
International publication number WO 2018016451
Date of international filing Jul 14, 2017
Date of international publication Jan 25, 2018
Priority data
  • P2016-144861 (Jul 22, 2016) JP
Title BROAD-SPECTRUM METHOD FOR DETERMINING E. COLI FLAGELLAR ANTIGEN TYPE USING PCR METHOD
Abstract [Problem] To provide a practical determination method whereby H type determination can be rapidly and easily performed.
[Solution] An Escherichia coli H typing determination method characterized by comprising determining H typing by performing nucleic acid amplification of a polymorphic region of a gene that codes for flagellin constituting flagella of E. coli. Specifically, through the present invention, the H type of E. coli can be genetically determined rapidly and accurately, and it is therefore possible to more appropriately and rapidly prevent infection with or infectious spread of enteropathogenic E. coli including enterohemorrhaghic E. coli in medical or public health/food sanitation settings.
Outline of related art and contending technology BACKGROUND ART
E. coli infection by pathogenic bacteria as well as the case occurs, the path a source of contamination and contamination, contamination as much as possible and quickly identified, it is necessary to prevent expansion of the infection.For this purpose, the characteristics of the isolates and correctly determined, the relationship between the strains needs to be clarified.
E. coli in the fine, another O-H based on a type and type-specific sera (O: H serum type) is, as a method for international gold standard are used for a long time.And another O-type, the E. coli bacteria present on the surface of the sugar chain structure as a method of classification as an antigen.In addition, H is a type, the motion of the organ as the flagellar antigen E. coli as a method of classification.The current, 'World Health Organization E. coli for study center in cooperation with a reference' Danish National Laboratory in serum (SSI: by Statens Serum Institut), as the fine E. coli, two types of O of the group 185 (O1-O188, subtype 3 type: O18ab/O18ac,O28ab/O28ac,O112ab,O112ac include, the type 6: O31,O47,O67,O72,O94,O122 is a vacant) and the 53 types of H-type (H1-H56,3 type: H13, H22, H50 is a vacant) is determined (non-patent document 1).
E. coli infection in the present state of the country, a serious cause severe (about 4,000 lines per year) is E. coli, Institute National Institute of infection by a local, substantially all of the isolates serotype determination is performed.However, the reagent is an anti-serum domestic manufacturer sells, corresponding to a part of not only serotype (O of the group: 50 type, H-type: type 22), and the determination cannot be made part of the separation lines has been a problem.A group of inventors, in order to solve this problem, almost all of the group of O can be determined genetically (E. coli O-genotyping PCR PCR method: ECOG-PCR) is developed and, in actual use in the inspection site and Patent Application are reached (Patent Document 1, Non-Patent Document 2).
However, O is determined by only a group of serum, it is not necessarily sufficient. For example, intestinal bleeding as E. coli is widely known in Japan and O157 is, as the name implies, in E. coli O157, Escherichia coli O157 in H7 is found in addition to the serious, pathogenic E. coli H45 or H39 of the gastrointestinal tract, such as a non-pathogenic E. coli H16, a plurality of H-type is known to be included.In addition, a serious national report a level higher than the number of Escherichia coli O103 is always separated, H2, H11, H25 such as H-type is included in the plurality of types.From these, as well as serum group O, H type can also be manufactured, another determination is performed by a precise mold is required.
Scope of claims (In Japanese)請求の範囲
[請求項1]
大腸菌の鞭毛を構成するフラジェリンをコードする遺伝子の多様性領域の核酸増幅を行うことにより,H型別の判定を行うことを特徴とする大腸菌H型別判定方法
[請求項2]
前記遺伝子が,fliC,flkA,fllA,flmAのいずれか又は複数から選択されることを特徴とする請求項1に記載の大腸菌H型別判定方法
[請求項3]
前記多様性領域が,配列番号1から53で示される配列のそれぞれにおいて,5’から500塩基,3’から300塩基を差し引いた配列領域から選択されることを特徴とする請求項1又は2に記載の大腸菌H型別判定方法
[請求項4]
前記多様性領域が,配列番号54から106で示される配列領域を含むことを特徴とする請求項3に記載の大腸菌H型別判定方法
[請求項5]
多様性領域の核酸増幅が,PCR法によりなされることを特徴とする請求項1から4に記載の大腸菌H型別判定方法
[請求項6]
核酸増幅が,プライマーセット1から51のいずれか又は複数のプライマーセットを用いてなされることを特徴とする請求項5に記載の大腸菌H型別判定方法
[請求項7]
請求項1から6に記載の大腸菌H型別判定方法に用いられることを特徴とするプライマー
[請求項8]
請求項1から6に記載の大腸菌H型別判定方法に用いられることを特徴とするプライマーセット
[請求項9]
請求項8に記載のプライマーセットを含み,大腸菌H型別判定方法に用いられるキット
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • UNIVERSITY OF MIYAZAKI
  • Inventor
  • IGUCHI Atsushi
  • BANJOU Masaya
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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