艾伯特埃希菌多重PCR检测方法的建立

Establishment of multiplex polymerase chain reaction assay for detection of Escherichia albertii

目的建立一种快速检测艾伯特埃希菌的多重PCR检测方法。方法根据lysP和EA0134基因建立多重PCR反应体系,并对引物浓度、反应时间及温度等反应条件进行优化,同时评价其灵敏性和特异性。运用该方法对实验室分离的63株艾伯特埃希菌菌株及增菌液和非艾伯特增菌液进行了检测。结果两对引物均能特异性扩增相应的基因,其灵敏性均为25 CFU/反应(500 CFU/ml)且非常特异。对本实验分离的艾伯特埃希菌增菌液和非艾伯特埃希菌增菌液的检测结果与之前传统方法的一致。结论本研究建立的多重PCR方法可用于艾伯特埃希菌的初步筛查和分离株的鉴定,为艾伯特埃希菌的快速鉴定和鉴别提供了一种新的检测方法。

Objective To develop a muhiplex polymerase chain reaction （PCR） assay for rapid identification of Escherichia albertii. Methods The multiplex PCR system was established based on lysP and EA0134 genes. Through optimizing the primer concentration, reaction time and temperature and other reaction conditions, the multiplex PCR system was evaluated by using 63 strains of Eseherichia albertii, Escherichia albertii and non-Escherichia albertii enrichment broth. Results The 2 pairs of primers could specifically amplify the corresponding genes, and the sensitivity was both 25 CFU/reaction （500 CFU/ml） and very specific. The detection results of enrichment broth of Escherichia albertii and non-Escherichia albertii isolated in this experiment were consistent with those previous conclusions drawn by the conventional assay. Conclusions The multiplex PCR system established in this study can be used to preliminary screen Escherichia albertii and identify the isolates, and this study provides an effective technological method for rapid identification of Escherichia albertii.