| 3种食源性致病菌Tem-PCR检测方法的建立 |
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| 引用本文： | 刘忠梅,徐义刚,曲敏,莫春生,刘新亮,李苏龙.3种食源性致病菌Tem-PCR检测方法的建立[J].食品科学,2016,37(4):186-190. |
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| 作者姓名： | 刘忠梅 徐义刚 曲敏 莫春生 刘新亮 李苏龙 |
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| 作者单位： | 1.黑龙江出入境检验检疫局，黑龙江 哈尔滨 150001；2.哈尔滨商业大学食品工程学院，黑龙江 哈尔滨 150076 |
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| 基金项目： | 公益性行业(质检)科研专项(201310126) |
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| 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。
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| 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 |
Development of Target-Enriched Multiplex PCR Assay for Simultaneous Detection of Three Foodborne Pathogens |
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| LIU Zhongmei;XU Yigang;QU Min;MO Chunsheng;LIU Xinliang;LI Sulong.Development of Target-Enriched Multiplex PCR Assay for Simultaneous Detection of Three Foodborne Pathogens[J].Food Science,2016,37(4):186-190. |
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| Authors: | LIU Zhongmei;XU Yigang;QU Min;MO Chunsheng;LIU Xinliang;LI Sulong |
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| Affiliation: | 1. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbin 150001, China;
2. College of Food Engineering, Harbin University of Commerce, Harbin 150076, China |
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| Abstract: | 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 femA gene, the Salmonella invA gene and
the Shigella ipaH gene, three pairs of specific primers together with a universal primer were designed. Following the
optimization of reaction conditions, the Tem-PCR assay was successfully established. Results showed that the Tem-
PCR assay was highly specific to the target bacteria with a sensitivity of 1.1 × 103 CFU/mL, 2 × 103 CFU/mL and
1.2 × 103 CFU/mL for Salmonella, S. aureus and Shigella, respectively. The Tem-PCR assay can effectively improve the
balance of amplification efficiencies of the primers employed in traditional multiplex PCR. |
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| Keywords: | Tem-PCR assay Salmonella Staphylococcus aureus Shigella |
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