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目的研究出一套高效、实用的水产品中霍乱弧菌检测方法,监测外环境和食品中霍乱弧菌污染状况,及时、有效地防控霍乱疫情。方法按照2012年版《全国霍乱监测方案》和第6版《霍乱防治手册》,配制含不同浓度霍乱弧菌的阳性水产品样品和不含霍乱弧菌的阴性水产品样品,用碱性蛋白胨水增菌6 h后,分别用胶体金法、上转发光法、实时荧光PCR法、分离培养法4种方法检测不同水产品中霍乱弧菌的检测效果。结果胶体金法、上转发光法、实时荧光PCR法、分离培养法检测水产品中霍乱弧菌,每种方法的检测时间和检测成本各不相同,方法的敏感性不同,但特异性无差异。结论在进行水产品中霍乱弧菌监测时,可以将4种方法有机结合,先选择1种~2种快速方法进行初筛。其中上转发光法是比较理想的快速筛查方法,对筛出的阳性标本再用分离培养法进一步确证和溯源分析,可以大大提高霍乱弧菌的检测效果,更加有效地防控霍乱疫情。
Objective To develop an efficient and practical method for the detection of Vibrio cholerae in aquatic products, monitor the contamination of Vibrio cholerae in the external environment and food, and prevent and control the outbreak of cholera in time and effectively. Methods According to the 2012 National Cholera Surveillance Program and the 6th Cholera Prevention and Treatment Manual, positive aquatic samples containing different concentrations of V. cholerae and negative aquatic samples not containing V. cholera were prepared and treated with alkaline peptone water After 6 hours, the detection methods of Vibrio cholerae in different aquatic products were tested by colloidal gold method, up-conversion luminescence method, real-time fluorescence PCR method and isolation culture method respectively. Results Colloidal gold method, up-conversion luminescence method, real-time fluorescence PCR method and isolation culture method were used to detect Vibrio cholerae in aquatic products. The detection time and detection cost of each method were different. The sensitivity of the method was different, but the specificity was no difference . Conclusion During the monitoring of Vibrio cholerae in aquatic products, four methods can be organically combined, and one or two rapid methods are selected first for screening. The up-conversion luminescence method is an ideal rapid screening method, and the positive samples that are screened out are further confirmed and analyzed by isolation culture method, which can greatly improve the detection effect of Vibrio cholerae and prevent and control the epidemic of cholera more effectively.