常见病原菌基因组DNA快速提取试剂盒研制

Preparation of rapid extraction kit of genomic DNA from common pathogens

以单增李斯特菌、金黄色葡萄球菌等6种常见食源性致病菌为研究对象，优化试剂盒工艺参数，采用琼脂糖凝胶电泳及OD260/OD280分析基因组DNA提取效果。结果表明，试验试剂盒可在30 min内完成基因组DNA提取，对常见10类食品样品抗基质干扰能力强。与实时荧光PCR技术联用，检测染菌量1～10 cfu·25 g-1食品样品时，仅一次增菌可得阳性结果；市售食品样品检测与国标方法结果无显著差异。所研制病原菌基因组DNA提取试剂盒适用于食品病原菌快速检测。

This study analyzed extraction quality of genomic DNA by agarose gel electrophoresis and OD260/OD280 to optimize technology parameters of kit, focusing on six kinds of common food-borne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, etc. The result showed that the extraction of genomic DNA could be finished in 30 min by the developed kit, and its anti-jamming capability was also strong in 10 categories of common food matrixes. The detection limits of the six kinds of pathogens could reach 1-10 cfu·25 g-1 samples by one enrichment culture based on real-time PCR. The detection results of commercial samples by the developed kit were consistent with GB detection. It can be used for rapid detection of pathogens in food.