快速提取细菌DNA方法的研究

目的:研究快速、有效可以应用于压电基因传感器对病原微生物的快速检测的DNA的提取方法。方法:采用沸水浴法,酚、氯仿法,chelex1 0 0法和试剂盒法提取金黄色葡萄球菌ATCC2 5 92 3、铜绿假单胞菌ATCC 2 785 3和大肠杆菌ATCC 2 5 92 2的DNA对产物进行纯度和产量测定,并对1 6SrDNA区域利用通用引物进行扩增。初步应用自行研制的压电基因传感器检测金黄色葡萄球菌ATCC2 5 92 3的PCR产物。结果:除沸水浴法,其他3种方法均能抽提出3种细菌的DNA与1 0 0法和酚、氯仿法抽提产物的纯度和产量差异无显著性(P >0 .0 5 ) ,但chelex1 0 0法操作更为简单、成本低;试剂盒法抽提出的产物纯度和产量明显高于酚、氯仿法、chelex1 0 0法P <0 . 0 5 ) ,但成本高,操作繁琐。结论:chelex1 0 0法操作简便、省时、成本低,适宜用于压电传感器样本的预处理。

THE RESEARCH OF RAPID DNA EXTRACTION FROM BACTERIA

Objective:To make comparison between four methods of DNA extraction from bacteria and select the best one for the detection of piezoelectric gene sensorMethods:DNA from Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853 and Ecoli ATCC 25922 was extracted through four methods including immersion method ,phenol/chloroform method,chelex100 method and EZNA kitThe purity and output of the products were detected and the frepuency shift of the piezoelectric gene sensor detection was observedResults:The mehtods except immersion method could extract DNA from the three strains of bacteriaThere was no significant difference between phenol/chloroform method and chelex100 method in terms of the purity and output of the products(P>005),but the chelex100 method is simpler and cheaperThe purity and output of DNA extracted by EZNA kit were obviously higher(P<005),but the method was more amplicated and expensiveConclusion:The chelex100 method is a simple,rapid and cheap method for DNA Extraction from bacteria and can be adopted in the pretreatment of piezoelectric gene sensor detection