实时定量PCR法快速检测水产品中的副溶血性弧菌  

Detection of Vibrio parahaemolyticus in seafood products by real-time quantitative PCR assay

作　　者：陈琳[1,2] 周青青[1] 顾青 郦萍[1] CHEN Lin;ZHOU Qingqing;GU Qing;LI Ping(Key Laboratory for Food Microbial Technology of Zhejiang Province, Zhejiang Gongshang University, Hangzhou 310018 , China;Hangzhou Wahaha Group Co., Ltd., Hangzhou 310018 , China)

机构地区：[1]浙江工商大学浙江省食品微生物技术研究重点实验室,浙江杭州310018 [2]杭州娃哈哈集团有限公司,浙江杭州310018

出　　处：《浙江农业学报》2019年第5期823-828,共6页Acta Agriculturae Zhejiangensis

基　　金：浙江省重大专项(2015C02039;2015C02022);国家自然科学基金(31606449;31871775);浙江省自然科学基金(LY16C200002;LQ18C200004)

摘　　要：副溶血性弧菌是一种广泛存在于水产品中的食源性致病菌,可污染食物,引起严重的食物中毒。利用实时定量PCR方法建立一种快速检测水产品中副溶血性弧菌的方法,根据随机扩增多态性分析(RAPD)鉴定特异性片段,设计特异性引物。采用建立的实时定量PCR方法检测市售水产品20份,检出阳性食品6份,最小检测灵敏度为50 fg DNA,与传统微生物检测结果相比无显著差异。该检测方法特异性强,灵敏度高。Vibrio parahaemolyticus is a food-borne pathogen, which can contaminate food and cause serious and acute gastroenteritis to human beings. In this study, a real-time polymerase chain reaction (PCR) assay was developed for the quantitative detection of Vibrio parahaemolyticus in aquatic products. Randomly amplified polymorphic DNA (RAPD) method was introduced to characterize species-specific DNA fragment of V. parahaemolyticus , and specific primers were designed. The proposed method was used to detect V. parahaemolyticus in 20 aquatic products on sale. It was shown that 6 samples out of 20 were carrying V. parahaemolyticus . The minimum detection sensitivity was 50 fg DNA. Compared with the methods recommended in national standard, the proposed method was swifter and more convenient to detect V. parahaemolyticus in auqatic products.

关 键 词：副溶血性弧菌 随机扩增多态性分析 实时荧光定量PCR 

分 类 号：S917.1[农业科学—水产科学] TS254.7[轻工技术与工程—水产品加工及贮藏工程]