3种方法对临床菌血症样本细菌DNA提取的比较  

Comparison of Three Methods for Extraction of Bacterial DNA from Bacteriemia Blood Samples

作　　者：赵晓丽[1] 李晓霞[1] 刘德华[1] 任宝军[1] 邵剑春[1] 胡大春[1] 

机构地区：[1]昆明市临床疾病分子生物学重点实验室,昆明医科大学附属甘美医院检验科,云南昆明650011

出　　处：《昆明医科大学学报》2014年第4期117-120,共4页Journal of Kunming Medical University

摘　　要：目的寻找一种简单、有效的提取临床菌血症样本细菌DNA的方法.方法分别用碱液加热裂解法、溶菌酶法和商品DNA提取液法提取细菌DNA,用临床常见病原菌特异性引物,进行PCR扩增.结果 (1)商品DNA提取液法提取革兰阴性菌DNA,经PCR扩增后得到特异性目的片段,但未扩增出革兰阳性菌的特异性目的片段;(2)碱液加热裂解法、溶菌酶法提取革兰阴、阳性菌DNA,经PCR扩增后均得到特异性目的片段,但碱液加热裂解法比溶菌酶法能扩增到更低浓度的细菌DNA目的片段.结论碱液加热裂解法是3种方法中最简单有效的细菌DNA提取方法,适用于临床病原菌基因组DNA提取.Objective To find an effective method for extracting bacterial DNA from bacteriemia blood samples. Methods Bacterial DNA from bacteriemia blood samples was extracted by three methods including alkali /heat lysis, lysozyme digestion and commercial DNA extraction kit. PCR with the specific primers for common pathogenic bacteria was used to amplify the extracted DNA. Result The PCR results showed that the methods of alkali/heat lysis was better than other two methods for extracting DNA from both Gram-negative and Gram-positive bacteria. Conclusion Alkali/heat lysis method is the most simple and effective method for extracting DNA from clinical common pathogenic bacteria.

关 键 词：细菌DNA 碱液加热裂解法 溶菌酶法 商品DNA提取法 

分 类 号：R378.99[医药卫生—病原生物学]