原料乳中多种致病菌的快速过滤富集及多重PCR检测  

Multiplex PCR Detection by Rapid Obtaining Foodborne Pathogens in Raw Milk with Filtration Method

作　　者：吕琦[1] 赵凤[1] 毕宇涵[1] 张光辉[1] 周艳秋[1] 遇晓杰[3] 张剑峰[3] 姜毓君[1,2] 

机构地区：[1]乳品科学教育部重点实验室,东北农业大学食品学院,黑龙江哈尔滨150030 [2]国家乳业工程技术研究中心,黑龙江哈尔滨150086 [3]黑龙江省疾病预防控制中心,黑龙江哈尔滨150030

出　　处：《食品与发酵工业》2008年第12期155-159,共5页Food and Fermentation Industries

基　　金：国家科技支撑计划(2006BAD04A08-13);教育部科学技术研究重点项目(208036);黑龙江省科技攻关项目(GA06C101-08);黑龙江省科技攻关项目(GC04C42501)

摘　　要：针对目前原料乳中致病菌的检测方法繁琐费时耗力的缺点,建立一种能同时检测原料乳中多种致病菌的快速检测方法,采用多重PCR快速灵敏检测技术与一种不需要预增菌就可以直接从原料乳中快速过滤富集菌体的技术相结合来同时检测原料乳中主要致病菌——沙门氏菌、大肠杆菌O157:H7、金黄色葡萄球菌、单增李斯特氏菌和蜡样芽孢杆菌。整个过程只需7～8h即可完成,且检测灵敏度可达102cfu/mL。这种方法是对传统检测方法的有效改进,并且达到了原料乳检测快速、准确、灵敏的要求。Conventional methods of detecting food borne pathogens in food are cumbersome and time-consuming. To establish a time-saving detecting method, pathogen were concentrated from raw milk by straight- forward filtration and were detected by a rapid and sensitive multiplex PCR system for simultaneous detection of Bacillus cereus, Salmonella spp, Escherichia coli O157：H7, Staphylococcus aureus and Listeria monocytogenes. The assay from filtration to multiplex PCR can be completed in about 7-8h and the limit of detection for multiplex PCR was as low as 102 cfu of bacteria in 1 mL of raw-milk. The study, which increases the accuracy and sensitivity, improves traditional methods and it can be applied to the food microorganism detection and diagnosis of food poisoning.

关 键 词：多重PCR 原料乳 致病菌 过滤 快速检测 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]