O1群和O139群霍乱弧菌荧光PCR快速检测方法的建立  

Establishment of a method for rapid detection of group O1 and O139 of Vibrio cholera by fluorescence PCR

作　　者：罗可天[1] 梁晅[1] 宋妙芳[1] 丁珊[1] 林智[1] 

机构地区：[1]广州市越秀区疾病预防控制中心,广东广州510055

出　　处：《中国热带医学》2009年第8期1402-1403,共2页China Tropical Medicine

基　　金：广州市越秀区科技局科技计划立项研究项目(2007-WS-006)

摘　　要：目的建立检测O1群和O139群霍乱弧菌的实时荧光聚合酶链反应(荧光PCR)方法,并进行优化和评价。方法根据Ol群和O139群霍乱弧菌O抗原编码基因设计探针和引物,建立同时检测霍乱弧菌O1群和O139群的荧光PCR方法,并对体系中的引物、Mg2+、dNTP和Taq酶进行优化,然后对建立的方法进行特异性、灵敏性、重复性的评价,并进行224份河口水样本的检测。结果建立了检测O1群和O139群霍乱弧菌的双重荧光PCR方法,对非O1群和O139群霍乱弧菌无扩增反应,敏感度比常规分离培养高。结论以O抗原编码基因为目标检测片段建立了O1群和O139群霍乱弧菌双重荧光PCR检测方法,可用于疑似霍乱弧菌感染的样本常规分离前的快速筛查。Objective To establish a methods for rapid detection of group Oland 0139 of Vibrio cholera by fluorescence PCR. Methods DNA probe and primers were designed based on O antigen code gene of Vibrio.cholera O1 and O139,and methods of simultaneous detection of group Oland O139 of Vibrio cholera by luorescent PCR was established and the primer,Mg^2＋ ,dNTP and Taq polymerase of the system were optimized. The specificity,sensitivity and reproducibility were assessed. Results A fluorescence PCR technique was established for simultaneous detection of group O1and O139 of Vibrio cholera and the sensitivity of the technique was higher apparently than normal isolation and culture method. Candusion The fluorescentce PCR technique targated at detection of O antigen code gene of group O1and O139 of Vibrio cholera has been established that allowed rapid screening of suspected cholera samples before conventional isolation.

关 键 词：霍乱弧菌 荧光PCR 方法优化 

分 类 号：R378.3[医药卫生—病原生物学]