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FOOD SCIENCE
2016,37
(4)
186-190
3种食源性致病菌Tem-PCR检测方法的建立
作 者：
刘忠梅;徐义刚;曲敏;莫春生;刘新亮;李苏龙
单 位：
黑龙江出入境检验检疫局;哈尔滨商业大学食品工程学院
关键词：
靶序列富集多重PCR方法;沙门氏菌;金黄色葡萄球菌;志贺氏菌
摘 要：
应用靶序列富集多重聚合酶链式反应(t arget-enriched multiplex-polymerase chain reaction,Tem-PCR)技术建立金黄色葡萄球菌、沙门氏菌和志贺氏菌3种食源性致病菌的快速检测方法。分别以金黄色葡萄球菌fem A基因、沙门氏菌inv A基因和志贺氏菌ipa H基因为靶基因,设计3对特异性引物,并与通用引物连接组成Tem-PCR引物,通过反应条件优化,建立了Tem-PCR检测方法。结果显示,建立的Tem-PCR方法特异性强,对沙门氏菌、金黄色葡萄球菌和志贺氏菌的检测灵敏度分别为1.1×10~3、2×10~3 CFU/m L和1.2×10~3 CFU/m L。Tem-PCR检测方法有效地解决了传统多重PCR引物扩增效率不均衡的问题。
译 名：
Development of Target-Enriched Multiplex PCR Assay for Simultaneous Detection of Three Foodborne Pathogens
作 者：
LIU Zhongmei;XU Yigang;QU Min;MO Chunsheng;LIU Xinliang;LI Sulong;Heilongjiang Entry-Exit Inspection and Quarantine Bureau;College of Food Engineering,Harbin University of Commerce;
关键词：
Tem-PCR assay;;Salmonella;;Staphyloc occus aureus;;Shigella
摘 要：
In this study,a target-enriched multiplex PCR(Tem-PCR) assay for simultaneous detection of Salmonella,Staphylococcus aureus and Shigella was developed. Based on the S. aureus fem A gene,the Salmonella inv A gene and the Shigella ipa H gene,three pairs of specific primers together with a universal primer were designed. Following the optimization of reaction conditions,the Tem-PCR assay was successfu lly established. Results showed that the TemPCR assay was highly specific to the target bacteria with a sensitivity of 1.1×10~3 CFU/mL,2×10~3 CFU/mL and 1.2×10~3 CFU/mL for Salmonella,S. aureus and Shigella,respectively. The Tem-PCR assay can effectively improve the balanc e of amplification efficiencies of the primers e mployed in traditional multiplex PCR.
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