多重荧光定量PCR同时检测霍乱弧菌、副溶血性弧菌和创伤弧菌的方法研究

Establishment of multiplex real-time PCR assay for detection of Vibrio cholerae,Vibrio parahaemolyticus and Vibrio vulnificus

目的:利用多重荧光定量PCR技术检测霍乱弧菌、副溶血性弧菌、创伤弧菌。方法:以溶藻弧菌等17种相关试验菌株做特异性检测;并将标准菌株稀释成不同梯度,做灵敏度检测。结果:与传统的检测方法相比,该方法有很好的特异性且灵敏度高,检测时间短并可节省人力,具有快速方便等优点。结论:适用于样品中霍乱弧菌、副溶血性弧菌和创伤弧菌的快速检测。检测限分别为:霍乱弧菌为280 cfu/ml;副溶血性弧菌为:100 cfu/ml;创伤弧菌为:450 cfu/ml。

Objective：Rapid detection of Vibrio cholerae;Vibrio parahaemolyticus and Vibrio vulnificus were established by multiplex real-time PCR assay.Methods： Seventeen test strains such as Vibrio alginolyticus was tested by specific detection.The different grades of standard strain were obtained by diluting and sensitivity was detected.Results： Compared with conventional method,the multiplex real-time PCR assay was specific and sensitively.Conclusion： The multiplex real-time PCR assay was rapid and accurate,with the detection limit of 280 cfu/ml for Vibrio cholerae,100 cfu/ml for Vibrio parahaemolyticus and 450 cfu/ml for Vibrio vulnificus.