Evaluation of TaqMan PCR assay for detecting <i>Salmonella</i> in raw meat and shrimp

作     者：Kimura, B Kawasaki, S Fujii, T Kusunoki, J Itoh, T Flood, SJA 

作者机构：Tokyo Univ Fisheries Dept Food Sci & Technol Minato Ku Tokyo 1088477 Japan Tokyo Metropolitan Res Lab Publ Hlth Shinjuku Ku Tokyo 1690073 Japan PE Appl Biosyst Foster City CA USA 

出 版 物：《JOURNAL OF FOOD PROTECTION》 (食品保护杂志)

年 卷 期：1999年第62卷第4期

页      面：329-335页

核心收录：

学科分类：0832[工学-食品科学与工程（可授工学、农学学位）] 08[工学] 0836[工学-生物工程] 

主　　题：培养基 DNA 细菌/分离和提纯 十足目(甲壳纲)/微生物学 评价研究(主题) 粪便/微生物学 食品微生物学 肉/微生物学 聚合酶链反应/方法 沙门菌属/生长和发育 沙门菌属/分离和提纯 沙门菌感染/微生物学 鼠伤寒沙门菌/分离和提纯 敏感性与特异性 血清分型 Taq聚合酶 动物 人类 

摘      要：We evaluated the TaqMan Salmonella amplification/detection kit from PE Applied Biosystems, which uses a polymerase chain reaction (PCR) assay for rapid detection of Salmonella in food samples. This system uses the 5 nuclease activity of Tag DNA polymerase, which digests an internal fluorogenic probe to monitor the amplification of the target gene. The system s sensitivity and specificity were evaluated using 42 serotypes of 68 Salmonella strains isolated from fecal samples from patients in Tokyo, Japan, and 39 non-Salmonella strains in 22 genera. There were no false-negative or false-positive results. This PCR assay can detect 3 CFU per PCR tube of Salmonella in pure culture (120 CFU/ml of TSB culture). PCR signals were attenuated with artificially contaminated shrimp, but a similar detection limit was obtained. TaqMan s performance was tested with 100 meat and chicken samples purchased from stores in Tokyo. Overall, two of the DNA extraction protocols (the Chelex and EnviroAmp methods) worked equally well, with some exceptions. Of the 100 samples analyzed, 10 were positive for Salmonella with both conventional culture methods and the kit and 89 were negative with both. One sample was negative by the culture method but positive by the kit assay. These results indicate that TaqMan is a reliable and rapid method for Salmonella analysis in the food industry. With this system, food samples can be analyzed for Salmonella in less than 20 h.