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目的建立生鲜猪肉中大肠杆菌和沙门菌的双重PCR检测方法,并初步调查生鲜猪肉中大肠杆菌和沙门菌的污染情况。方法以大肠杆菌O157∶H7 rfb E基因和沙门菌inv A基因为靶基因设计引物。通过对单个基因PCR和多重基因PCR扩增进行特异性、敏感性试验以及优化反应体系,建立快速检测大肠杆菌和沙门菌的双重PCR法。在郑州市不同地区的综合性超市、冷鲜肉专卖店和农贸市场随机抽检144份生鲜猪肉样品,分别进行了PCR检测和常规微生物学检验。结果建立的双重PCR方法特异性好,抗干扰能力强,灵敏度可达到10 pg/μl。在144份样品中检测出大肠杆菌的样品数为10份,检出率为6.94%;检出沙门菌的样品数为13份,检出率为9.03%;大肠杆菌和沙门菌同时检出的有2份,占总样品的1.39%。结论初步建立了同步、简便、快速、灵敏地检测生鲜猪肉中大肠杆菌、沙门菌的双重PCR方法;生鲜猪肉中存在致病菌的污染问题,将威胁到食品安全和人体健康,不容忽视。
Objective To establish a dual PCR method for the detection of Escherichia coli and Salmonella in fresh pork and to investigate the contamination of Escherichia coli and Salmonella in fresh pork. Methods The primers of E. coli O157: H7 rfb E gene and Salmonella inv A gene were designed as target genes. By carrying out specific and sensitive experiments on single gene PCR and multiple gene PCR amplification and optimizing the reaction system, a dual PCR method for rapid detection of Escherichia coli and Salmonella was established. In different areas of Zhengzhou City, a comprehensive supermarket, cold meat stores and farmers market randomly sampled 144 samples of fresh pork were PCR and routine microbiological tests. The results of the double PCR method established good specificity, anti-interference ability, sensitivity up to 10 pg / μl. In 144 samples, E. coli was detected in 10 samples and the detection rate was 6.94%. The number of Salmonella samples was 13 and the detection rate was 9.03%. Simultaneous detection of Escherichia coli and Salmonella There are 2, accounting for 1.39% of the total sample. Conclusions A dual PCR method for the simultaneous, simple, rapid and sensitive detection of Escherichia coli and Salmonella in fresh pork has been initially established. The contamination of pathogenic bacteria in fresh pork is threatened by food safety and human health, which can not be ignored .