Detection of the Virulence-associated Genes in Vibrio Cholerae by Multiplex PCR Assay
- VernacularTitle:多重PCR检测霍乱弧菌的毒素相关基因
- Author:
Liangyi JING
;
Jinying CHEN
;
Shumei WANG
- Publication Type:Journal Article
- Keywords:
Vibrio Cholerae;
Polymerase chain reaction;
Virulence-associated genes
- From:
Journal of Environment and Health
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study a rapid and sensitive method for determination of virulence-associated genes in O1El Tor,O139,non-O1/non-O139Vibrio cholerae strains.Methods Five pairs of primers were designed respectively ac-cording to cholera toxin sub-unit A gene(ctxA),accessory cholera enterotoxin gene(ace),zonula occludens toxin gene(zot),toxin coregulated pilus A gene(tcpA)and toxR regulatory protein gene(toxR).Multiplex PCR(MPCR)procedures for simultaneously detecting those five genes were established.The gene information of the virulence-associated genes in the Vibrio cholerae strains was obtained through agar gel electrophoresis for products of single amplification of the MPCR.Results The five virulence-associated genes in the positive control Vibrio cholerae O139(MO45strain)could be detected and the results were correct,which could meet the designed request for the method.In the other tested strains(O1EL Tor,O139,non-O1/non-O139)could be detected1to5kinds of the virulence-associated genes.Based on the results of the variety of carried virulence-associated genes,the tested Vibrio cholerae strains could be classified as5kinds of genetypes,and the Vibrio cholerae could be distinguished between toxic and non-toxic strains.The sensitivity of the MPCR approach reached to10 2 cfu/ml.Conclusion This method is rapid,specific and sensitive,which possess great value for practical application.
