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目的:建立基于TaqMan探针的荧光定量PCR检测水产品中沙门菌的检测体系。方法:根据沙门菌外膜蛋白Ompc基因的保守序列,设计1对引物和TaqMan探针,并对荧光定量PCR反应体系和反应条件进行优化。利用建立的检测方法对舟山水产品加工厂和舟山菜市场采集的120份样本进行检测。结果:该检测方法可以达到1μl 1.0×101拷贝数量级的灵敏度。样品检出率为10.0%,与国标法检测结果一致。结论:本研究建立的TaqMan探针荧光定量PCR检测方法可以特异、灵敏、简便快速的实现对水产品中沙门菌的检测。
Objective: To establish a detection system of Salmonella in aquatic products by real-time fluorescent quantitative PCR based on TaqMan probe. Methods: According to the conserved sequence of Ompc gene of Salmonella outer membrane, a pair of primers and TaqMan probe were designed. The system of fluorescence quantitative PCR and reaction conditions were optimized. 120 samples collected from Zhoushan Aquatic Products Processing Factory and Zhoushan Vegetable Market were tested by the established test method. Results: This assay achieves a sensitivity of the order of 1 x 1.0 x 101 copies. The sample detection rate was 10.0%, consistent with the national standard test results. Conclusion: The TaqMan probe fluorescence quantitative PCR assay established in this study can detect Salmonella in aquatic products in a specific, sensitive, simple and rapid way.