适用于二代测序技术的细菌DNA提取方法的比较研究  

Comparative Study on Bacterial DNA Extraction Methods for NGS Technique

作　　者：陈佩[1,2] 岳巧云 刘德星 陈利伟 邱德义 何秋星 CHEN Pei;YUE Qiaoyun;LIU Dexing;CHEN Liwei;QIU Deyi;HE Qiuxing(Guangdong Pharmaceutical University,Guangzhou,Guangdong,510006,China;Zhongshan Entry-Exit Inspection and Quarantine Bureau)

机构地区：[1]广东药科大学,广东广州510006 [2]中山出入境检验检疫局

出　　处：《检验检疫学刊》2018年第2期55-60,29,共7页Journal of Inspection and Quarantine

基　　金：国家重点研发计划项目(2016YFF0203205);国家质量监督检验检疫总局科技计划项目(2017IK165);广东检验检疫局科技计划项目(2017GDK63)

摘　　要：为建立用于二代测序技术的最适细菌DNA提取方法。分别用磁珠法、热裂解法、试剂盒法(有溶菌酶处理和无溶菌酶处理)、超声波法、超声+热裂解法共6种方法提取等量混合的细菌DNA,特异性扩增其16S r RNA的V3-V4高变区基因,经二代测序并分析结果中的细菌种类及其相对含量分布。6种方法的测序结果中,细菌种类鉴定结果无明显差异;而在磁珠法和试剂盒法(有溶菌酶处理)结果中,不同细菌的含量分布最为接近样品中的真实情况。在6种方法中,磁珠法和试剂盒法(有溶菌酶处理)适合于二代测序技术中混合细菌DNA的提取。To establish the most suitable bacterial DNA extraction method for Second-generation sequencing technology. The genomic DNA from equal mixed bacteria was extracted by totally six different methods respectively,including magnetic method,pyrolysis method,DNA extraction kit method（with lysozyme treatment or without lysozyme treatment）,ultrasonic or ultrasonic combined with pyrolysis method. The DNA fragment of 16 S r RNA V3-V4 hypervariable region was amplified specifically. And then the bacterial species and their relative distribution were estimated according to the DNA sequencing analysis. There were no significant differences for the identification of bacterial species among the sequencing results from the six methods. However,the distribution of different bacterial contents was relatively uniform for the results from magnetic method and DNA extraction kit method（lysozyme treatment）. The magnetic method and DNA extraction kit method（lysozyme treatment） are suitable for the extraction of mixed bacterial DNA purification in Second-generation sequencing technology.

关 键 词：二代测序技术 细菌DNA提取方法 16S r RNA V3-V4高变区 

分 类 号：R37[医药卫生—病原生物学]