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目的 建立检测O1群和0139群霍乱孤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O1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PER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群霍乱孤菌的双重荧光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 O1 and O139 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 O1 and O139 of Vibrio cholera by luorescent PCR was established and the primer,Mg2+ ,dNTP and Taq polyroerase of the system were optimized. The specificity,sensitivity and reproducibility were assessed. Results A fluorescence PCR technique was established for simultaneous detection of group O1 and O139 of Vibrio cholera and the sensitivity of the technique was higher apparently than normal isolation and culture method. Conclusion The fluorescentce PCR technique targatod at detection of O antigen code gene of group O1 and O139 of Vibrio cholera has been established that allowed rapid screening of suspected cholera samples before conventional isolation..