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复合探针实时荧光PCR技术快速检测海产品中 副溶血性弧菌方法的建立
闻洁,陆利霞,熊晓辉
0
(南京工业大学食品与轻工学院,南京 210009;南京工业大学食品与轻工学院,南京 210009; 江苏省食品安全快速检测公共技术服务中心,南京 210009)
摘要:
副溶血性弧菌是一种食源性致病菌,海产品和其制品海鲜酱油等最容易被副溶血性弧菌污染,而副溶血性弧菌寄生于鲜虾、海鲜酱油等复杂的食品基质中有时却很难被检测出。以快速检测食品中的副溶血性弧菌为目的,针对副溶血性弧菌的tlh基因,设计引物和复合探针,采用实时荧光PCR方法检测鲜虾和海鲜酱油中的副溶血性弧菌,得出以下结论,该方法检测副溶血性弧菌具有较强的特异性和灵敏度,当鲜虾(固体)中的样品菌含量为103cfu·g-1或海鲜酱油中的样品菌含量为103cfu·mL-1时,无需增菌培养,即可快速检出。增菌6~8 h即可检出鲜虾(固体)或海鲜酱油(液体)的1cfu的副溶血性弧菌。
关键词:  副溶血性弧菌  聚合酶链式反应  复合探针  荧光
DOI:10.13610/j.cnki.1672-352x.20161205.021
基金项目:江苏省科技厅社会发展项目(BE2016803)资助。
Establishment of rapid detection of Vibrio parahaemolyticus in seafood by real-time fluorescent PCR with complex probe
WEN Jie,LU Lixia,XIONG Xiaohui
(College of Food Science and Light Industry, Nanjing University of Technology, Nanjing 210009;College of Food Science and Light Industry, Nanjing University of Technology, Nanjing 210009; Jiangsu Public Technical Service Center for Rapid Detection of Food Safety, Nanjing 210009)
Abstract:
Vibrio parahaemolyticus is a kind of food-borne pathogens, which is easily caught in seafood. Shrimp is rich in protein and fat, carbohydrates, calcium, phosphorus, iron and other ingredients. The ingredients of seafood sauce is complex, containing scallops, scallop powder, spices, rice wine besides salt and monosodium glutamate. It is difficult to be detected in such complex food matrix. In this paper, primers and complex probe are designed, based on the tlh gene ,to detect Vibrio parahaemolyticus in shrimp and seafood sauce by real-time PCR. The results are as follows: It can be quickly checked out without culture under the bacteria content of 103 cfu whether in shrimp samples (solid) or in seafood soy sauce samples. It can be detected by fluorescent PCR with the enrichment of 6-8 h in the shrimp (solid) or seafood sauce (liquid).
Key words:  Vibrio parahaemolyticus  PCR(polymerase chain reaction)  complex probe  fluorescence

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