文献类型：期刊文献

中文题名：四种桉树青枯菌DNA提取方法及PCR检测灵敏度比较

英文题名：Comparison of four different DNA extraction methods of Ralstonia solanacearum in eucalyptus and sensitivity of PCR detection

作者：王胜坤[1] 王军[2] 徐大平[1]

第一作者：王胜坤

机构：[1]中国林业科学研究院热带林业研究所;[2]华南农业大学林学院

年份：2007

卷号：26

期号：5

起止页码：4-7

中文期刊名：中国森林病虫

外文期刊名：Forest Pest and Disease

收录：CSTPCD;;北大核心:【北大核心2004】；

基金：国家科技攻关项目"南方主要速生阔叶树种新品种选育及栽培技术"(2004BA515B02);科技部成果转化项目"南方主要速生阔叶树种新品种及高效培育技术中试"

语种：中文

中文关键词：青枯菌;DNA提取;PCR检测;灵敏度

外文关键词：Ralstonia solanacearum ; DNA extraction; PCR detection; sensitivity

分类号：S763.15

摘要：以带有青枯菌Ralstonia solanacearum的桉树组织为材料,采用4种不同的提取方法抽提青枯菌DNA,同时利用青枯菌的特异性引物进行PCR扩增,比较了4种DNA的提取方法及PCR检测的灵敏度。结果表明,煮沸法和热裂解法操作相对简单,但检测灵敏度较低,检测限分别为104CFU/mL和103CFU/mL;简化提取法和试剂盒法检测效果较好,均可检测到102CFU/mL的青枯菌;简化提取法比试剂盒法操作更简单,检测成本低,具有更高的应用价值。
The DNA of Ralstonia solanacearum was extracted from eucalyptus tissue with four different methods and amplified with specific primers of R. solanacearum. The sensitivities of PCR detection were compared. The results showed that the methods of boiling and cell lysis by heating were easier to operate but had lower sensitivities with 10^4 CFU/mL and 10^3 CFU/mL, respectively. DNA-extraction kit method and simple extraction method had better detection effects, The detection limit of both methods was 10^2 CFU/mL. The simple extraction method with the advantage of easier operation and lower cost may have higher application value.