使用实时聚合酶链反应快速检测食品中的沙门氏菌。

Rapid detection of Salmonella in foods using real-time PCR.

作者信息

Cheng Chorng-Ming, Lin Wen, Van Khanh Thien, Phan Lieuchi, Tran Nelly N, Farmer Doris

机构信息

U.S. Food and Drug Administration, Pacific Regional Laboratory Southwest, Irvine, California 92612, USA.

出版信息

J Food Prot. 2008 Dec;71(12):2436-41. doi: 10.4315/0362-028x-71.12.2436.

DOI:10.4315/0362-028x-71.12.2436

PMID:19256088

Abstract

Conventional methods for detection of Salmonella serovars in foods are generally time-consuming and labor intensive. A real-time PCR method has been developed with custom designed primers and a TaqMan probe to detect the presence of a 262-bp fragment of the Salmonella-specific invA gene. The method has been tested with a total of 384 field-isolated Salmonella serovars and non-Salmonella stock strains, as well as 420 U.S. Food and Drug Administration food samples, comprising a variety of food matrices. The method was highly specific in detecting Salmonella in spiked chili powder and shrimp samples, with a sensitivity of 0.04 CFU/g. In addition, the method is faster, more accurate, and less costly than the traditional U.S. Food and Drug Administration's Bacteriological Analytical Manual cell-culturing and the AOAC International-approved VIDAS methods to detect Salmonella in foods.

摘要

食品中沙门氏菌血清型的传统检测方法通常耗时且 labor intensive。已开发出一种实时聚合酶链反应（PCR）方法，使用定制设计的引物和 TaqMan 探针来检测沙门氏菌特异性 invA 基因 262 碱基对片段的存在。该方法已用总共 384 株现场分离的沙门氏菌血清型和非沙门氏菌标准菌株以及 420 份美国食品药品监督管理局的食品样本进行了测试，这些样本包含多种食品基质。该方法在检测加标辣椒粉和虾样本中的沙门氏菌时具有高度特异性，灵敏度为 0.04 CFU/g。此外，与美国食品药品监督管理局传统的《细菌学分析手册》细胞培养法以及美国官方分析化学师协会（AOAC International）批准的 VIDAS 方法相比，该方法检测食品中沙门氏菌的速度更快、更准确且成本更低。