多重聚合酶链式反应(MPCR)用于霍乱弧菌检测的研究  

Detection of Vibrio cholerae by multiplex polymerase chain reaction(MPCR)

作　　者：刘彩莲[1,2] 祁国明[1,2] 徐建国[1,2] 刘延清[1,2] 高守一 

机构地区：[1]中国预防医学科学院流行病学微生物学研究所 [2]卫生部医学分子细菌学重点实验室

出　　处：《中华微生物学和免疫学杂志》1997年第4期314-319,共6页Chinese Journal of Microbiology and Immunology

基　　金：卫生部科研基金

摘　　要：本文首先从实用的角度探讨用三对特异的引物对不同血清群霍乱弧菌进行ＭＰＣＲ检测的可能性。这三对引物具有很大的应用价值，从一次扩增反应中可以得出有关菌株的毒力因子和血清群的信息。检测的敏感性可达到１０ＣＦＵ。实验还提示将引物数量改为五条寡核苷酸链来进行ＭＰＣＲ能得出同样的结果。对Ｐ３、Ｐ４引物扩增形成的６１７ｂｐ的ＤＮＡ片段进行酶切分析表明，埃尔托型霍乱弧菌Ｏ１３９群霍乱弧菌的ｔｃｐＡ基因有很大的相似性，至少在ＰｓｔⅠ和ＢｇｌⅡ酶切位点上表现相同，但两者均与古典型霍乱弧菌不同。From practival point of view,the paper firstly discussed the possibility to detect different serogroups of Vibrio cholerae with MPCR using three pairs of specific primers.These three pairs of specific primers possessed great value for practical application.We could obtain virulent factor and serogroup information of related strains through single amplification.The sensitivity of MPCR reached 10 CFU.Through experiments,it indicated that the same results would be obtained using five oligonucleotide chains instead of the three pairs of primers.With restriction endonuclease to analyse DNA fragments of 617bp formed by P3 and P4 primers amplification,we claimed,great resemblance exists in tcpA gene of E1 Tor V.cholerae and V.cholerae O139,at least at the sites of PstⅠ and BglⅡ digestions,but different in both sites from classical V.cholerae .

关 键 词：聚合酶链反应 霍乱弧菌 毒素基因 检测 

分 类 号：R516.504[医药卫生—内科学]