环介导等温扩增
金黄色葡萄球菌
产肠毒素大肠杆菌
DNA提取
微生物学
聚合酶链反应
基因组DNA
食品科学
生物
DNA
色谱法
化学
细菌
基因
大肠杆菌
肠毒素
遗传学
作者
N Sowmya,M.S. Thakur,H.K. Manonmani
标识
DOI:10.1111/j.1365-2672.2012.05315.x
摘要
The study describes the development of simple and rapid DNA extraction method in combination with loop-mediated isothermal amplification (LAMP) to detect enterotoxigenic Staphylococcus aureus in food samples. In this study, isolation of genomic DNA of enterotoxigenic Staph. aureus from spiked milk, milk burfi, khoa, sugarcane juice and boiled rice was carried out by boiling the isolated sample pellets for 10 min with 1% Triton X-100. The isolated DNA was evaluated by polymerase chain reaction (PCR) and LAMP method. The LAMP was found to be 100 times more sensitive than PCR. The LAMP assay was very specific for Staph. aureus, and the presence of other contaminating bacterial DNAs and food matrix did not interfere or inhibit the LAMP assay. The template DNA extraction method developed in this study for food samples is simple, rapid and cost-effective. LAMP was found to be less sensitive to matrix effect of food, compared to PCR. The method is suitable for direct detection of Staph. aureus without any enrichment in contaminated food samples and hence finds its application in food safety analysis, in permutation with LAMP.
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