速冻面米制品中金黄色葡萄球菌DNA提取方法的比较分析
首发时间:2016-05-23
摘要:DNA的提取效率是影响食品中致病菌PCR检测的关键因素。因此本研究以速冻面米制品中最易污染的金黄色葡萄球菌为研究对象,比较了四种DNA提取方法(CTAB法、离心柱法、氨基化纳米磁珠法、羧基化纳米磁珠法)的效率和质量,对提取的DNA采用Real-time PCR方法进行评价。结果表明,在三大类速冻面米制品(肉馅类、素馅类和无馅类)人工污染金黄色葡萄球菌的样品中,羧基化纳米磁珠法和氨基化羧纳米磁珠法的提取效率显著优于CTAB法和离心柱法,可推荐为速冻面米制品中金黄色葡萄球菌污染快速定量检测的DNA提取方法。
关键词: 食品卫生与检验 金黄色葡萄球菌 Real-time PCR 速冻面米制品 DNA提取
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Comparison on DNA Extraction Methods of Staphylococcus aureus from Quick-Frozen Flour and Rice Products
Abstract:DNA extraction is the critical step in the process of detecting pathogens in food by PCR. In this study, we compared four DNA extraction methods of Staphylococcus aureus from artificially contaminated quick-frozen flour and rice products. Genomic DNA extraction was performed using the CTAB method, spin column kit method, amino-modified silica-coated magnetic nanoparticles method and carboxyl beads kit method, then the four methods were evaluated by Real-time PCR. The results showed that silica-coated magnetic nanoparticles method and carboxyl beads kit method were significantly more effective than CTAB method and spin column kit method when extracting DNA from three different kinds of quick-frozen flour and rice products. So silica-coated magnetic nanoparticles method and carboxyl beads kit method can be cooperated with Real-time PCR in rapid quantitative detection of Staphylococcus aureus in quick-frozen flour and rice products.
Keywords: Food hygiene and inspection;Staphylococcus aureus Real-time PCR quick-frozen flour and rice products DNA extraction
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