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投稿时间：2018-08-20
投稿时间：2018-08-20
中文摘要: 建立快速检测餐饮食品中沙门氏菌、金黄色葡萄球菌、单核增生李斯特氏菌、大肠埃希氏菌O157：H7及副溶血性弧菌5种致病菌的实时聚合酶链式反应体系（real-time polymerase chain reaction，RT-PCR）。通过增菌时间的比较，调整RT-PCR反映体系中的退火温度，建立5种致病菌相同的检测体系。试验结果表明，5个目标菌达到检测灵敏度的最短增菌时间范围为2 h～8 h，5种致病菌的试验灵敏度范围为7.4×10 CFU/mL～2.4×103CFU/mL。5种目标菌的熔解温度Tm值分别为沙门氏菌87.32℃，金黄色葡萄球菌79.57℃，单核增生李斯特氏菌82.02℃，大肠埃希氏菌O157：H7 82.24℃，副溶血性弧菌87.15℃。试验建立的RT-PCR快速检测反应体系，与常规微生物检测方法相比，有效地缩短了检测时间，且灵敏度与特异性均能满足检测要求。
Abstract:Abstract：A method to detect Salmonella，Staphylococcus aure，Listeria monocytogenes，Escherichia coli O157：H7 and Vibrio Parahemolyticus in catering food by Real-time PCR with melting curve was established.Compared different enrich time of 5 pathogenic bacteria，in order to find the shortest enrich time of each bacteria.Adjusted the annealing temperature to find the same suitable PCR system.The method showed that the range of shortest enrich time of each pathogenic bacteria was from 2 h to 8 h，and the sensibility was ranged from 7.4×10 CFU/mL to 2.4×103CFU/mL.The TM values were 87.32，79.57，82.02，82.24℃ and 87.15℃.The rapid system of Real-time PCR with melting curve could effectively short the detecting time，and the sensibility and specificity were suitable for testing.
keywords: real-time polymerase chain reaction melting curve Salmonella Staphylococcus aureus Listeria monocytogenes Escherichia coli O157：H7 Vibrio Parahemolyticus
文章编号：201904027 中图分类号： 文献标志码：
基金项目:基金项目：嘉兴市科技计划项目（2017BY18070）
| Author Name | Affiliation |
| YAO Yan-ling，LIU Luo-qiang，GUAN Jia-li | Jiaxing Testing Institute for Food and Drug Control，Jiaxing 314050，Zhejiang，China |
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