Chelex-100法和碱性裂解法提取细菌DNA的比较  

Comparison of methods for extraction of DNA by chelex-100 and alkali split

作　　者：钱雪琴[1] 张军[1] 沈芳[1] 

机构地区：[1]上海市公共卫生中心,上海201508

出　　处：《中国卫生检验杂志》2008年第8期1565-1566,共2页Chinese Journal of Health Laboratory Technology

摘　　要：目的:通过对两种DNA提取方法的比较,寻找一种灵敏、快速、经济实用的提取方法。方法:采用Chelex-100法和碱性裂解法分别提取不同浓度大肠杆菌ATCC25922和金黄色葡萄球菌ATCC25923的DNA,利用通用引物扩增16SrDNA片段检测两种方法的灵敏度。结果:碱性裂解法提取的不同浓度的两种细菌DNA,经PCR扩增均未显出目的条带;Chelex-100法提取不同浓度的两种细菌DNA均扩增出目的条带,PCR检测大肠埃希菌和金黄色葡萄球菌的灵敏度均为1.2×101cfu/ml,电泳结果提示相同浓度的大肠杆菌被提取到的DNA含量比金黄色葡萄球菌的含量高。结论:Chelex-100法操作简便,可用于细菌DNA的快速提取。Objective： To find a sensitive,rapid and economic method of DNA extraction by comparison of Chelex-100 with alkali split.Methods：DNA from different concentrations of E.coli ATCC25922 and Staphylococcus aureus ATCC25923 was extracted using Chelex-100 and Alkali split methods.The extracted DNA was amplified using universal primers for 16SrDNA fragment.Results：No signal was obtained using PCR on the DNA samples extracted from different concentrations of two bacteria by alkali split.All DNA samples extracted from different concentrations of two bacteria by Chelex-100 were amplified.The sensitivity of PCR was 1.2×101 cfu/ml for either the detection of E.coli or Staphylococcus.The result of electrophoresis showed that the DNA levels were higher in E.coli than in Staphylococcus using the same concentration of bacteria for DNA extraction.Conclusion：The Chelex-100 extraction method is a simple and valuable method for rapid isolation of DNA from bacteria.

关 键 词：Chelex-100 PCR 16SrDNA 碱性裂解法 

分 类 号：R446.5[医药卫生—诊断学]