摘要
本研究对人工接种肠出血性大肠杆菌O157∶H7和肠侵袭性大肠杆菌的莴苣样品中菌体的富集和DNA提取方法进行了优化。研究中比较了不同过滤膜组合对菌体的富集效果,筛选了莴苣样品中细菌DNA提取的最适方法,建立了多重PCR方法检测人工污染莴苣样品。结果表明,采用尼龙膜15μm+混合膜0.22μm的组合富集细菌、试剂盒法提取样品中细菌DNA,多重PCR检测的检出限降低10倍,检测时间节省1.5 h。本研究可快速、简便、灵敏的应用于莴苣检测中,具有很高的实际应用价值。
To optimize the methods for concentration and DNA extraction of two pathogens in artificial lettuce infected with Enterohemorrhagic E.cofi and Enteroinvasive E.coli. This study compared the concentration effect of different kinds of filtration membranes,and selected the best method of DNA extraction to extract the two athogens in lettuce sample. The multiplex PCR was used to detect the artificial lettuce infected with two pathogens. The results showed that the nylon membrane which had the pore size of 15 μm combined with mixed membrane which had the pore size of 0.22 μm was the best enrichment method. And the best DNA extraction method was slected by using the kit of bacterial genomic DNA extraction. The detection limit of lettuce samples by using multiplex PCR was decreased 10 times. The detection time was saved 1.5 h. The multiplex PCR combined with the optimized method was sensitive,fast and easy. This study had a high practical value.
出处
《食品工业科技》
CAS
CSCD
北大核心
2015年第20期260-264,270,共6页
Science and Technology of Food Industry
基金
国家科技支撑计划项目(2012BAD38B05)
国家自然科学基金项目(31172009)
大连市科技计划项目(2012E13SF106)
大连市金州新区科技计划项目(2012-A1-049)
关键词
鲜活农产品
致泻大肠杆菌
富集
DNA提取
fresh agricultural products
diarrheagenic Escherichia cofi
bacteria concentration
DNA extraction
