多重荧光定量PCR法检测海产品中副溶血性弧菌、沙门菌和单增李斯特菌

发布时间：2018-03-21 23:38

本文选题：海产品　切入点：副溶血性弧菌　出处：《中国食品卫生杂志》2016年04期 　论文类型：期刊论文

【摘要】：目的建立检测海产品中副溶血性弧菌、沙门菌和单增李斯特菌的多重荧光定量PCR体系。方法针对副溶血性弧菌tlh基因,沙门菌Ompc基因和单增李斯特菌hly基因设计引物和Taq Man探针,建立多重荧光定量PCR体系,进行特异性与敏感性研究;利用该体系检测海产品中的副溶血性弧菌、沙门菌和单增李斯特菌。结果副溶血性弧菌、沙门菌和单增李斯特菌可得到特异性扩增,而共存于海产品中的其他细菌均未见扩增曲线。敏感性试验显示,该体系对副溶血性弧菌、沙门菌和单增李斯特菌的最低检测限分别为72、40、80 cfu/ml。对舟山采集的150份样品进行检测,检出32份副溶血性弧菌、11份沙门菌、5份单增李斯特菌,与国标法检测结果一致。结论本研究建立的基于Taq Man探针的多重荧光定量PCR检测方法可以特异、灵敏、简单快速地实现对海产品中副溶血性弧菌、沙门菌和单增李斯特菌的检测。
[Abstract]:Objective to establish a multiplex fluorescence quantitative PCR system for the detection of Vibrio parahaemolyticus Salmonella and Listeria monocytogenes in marine products. Ompc gene of Salmonella monocytogenes and hly gene of Listeria monocytogenes were designed with primers and Taq Man probes to establish a multiplex fluorescent quantitative PCR system to study the specificity and sensitivity of Vibrio parahaemolyticus in seafood. Results specific amplification of Vibrio parahaemolyticus, Salmonella and Listeria monocytogenes was obtained, but no expansion curve was found in other bacteria coexisting in seafood. The minimum detection limit for Vibrio parahaemolyticus, Salmonella and Listeria monocytogenes was 724080cfur / ml, respectively. 150 samples collected from Zhoushan were detected, and 32 strains of Vibrio parahaemolyticus (Vibrio parahaemolyticus), 11 samples of Salmonella, 5 strains of Listeria monocytogenes were detected. Conclusion the multiplex fluorescence quantitative PCR method based on Taq Man probe can be used to detect Vibrio parahaemolyticus in marine products. Detection of Salmonella and Listeria monocytogenes.
【作者单位】：舟山出入境检验检疫局;浙江海洋学院;
【基金】：浙江省分析测试科技计划项目(2014C38108)
【分类号】：R155.5

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