FTA滤膜与多重PCR结合检测肉中3种食源性致病菌  

Detection of Three Food-borne Bacterial Pathogens in Meat Using FTA Filtering Combined with Multiplex PCR Assay

作　　者：杨国兴[1] 苏旭东[1] 李艾[2] 周巍[1] 张亚爽[1] 张伟[1] 

机构地区：[1]河北农业大学食品科技学院,河北保定071000 [2]唐山学院环境与化学工程系,河北唐山063000

出　　处：《中国食品学报》2009年第4期198-204,共7页Journal of Chinese Institute Of Food Science and Technology

摘　　要：采用FTA滤膜从肉中直接提取模板DNA,根据金黄色葡萄球菌的nuc基因、沙门氏菌的IpaB基因、福氏志贺氏菌的ipaH基因设计3对特异性引物,进行多重PCR检测。结果表明,3对引物能特异性扩增出210、280、393bp大小的目的条带;在不增菌的情况下,多重PCR同时检测肉中3种致病菌的灵敏度均为102cfu/g,检测时间6h。该检测方法准确、快速、高效,为同时检测肉中金黄色葡萄球菌、沙门氏菌、福氏志贺氏菌奠定了基础。A multiplex PCR was designed for the simultaneous detection of the Bacterial Pathogens （Staphylococcus aureus, Salmonella spp, and Shigellaflexneri） involved in meat. Primer targeting the nuc, ipaB and ipaH were used in the PCR. Each of the three pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. Based on flotation,solvent extraction technology, FTA filter was used to extract pathogens DNA from artifically contaminated meat. Three DNA fragments of 210bp, 280bp and 393 bp were amplified. The sensitivity of the multiplex PCR was 10^2 cfu/g for Staphylococcus aureus, Salmonella spp and Shigella flexneri in meat. The developed method in this assay allows detection of the pathogens in meat in less than 6 h. This method lies on its accuracy, rapidity and efficiency in the diagnosis, so it is a useful method for the simultaneous detection of the three species of bacteria in meat.

关 键 词：多重PCR 食源性致病菌 FTA滤膜 肉 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]