不同的DNA提取法对两种常见医院感染菌ERIC-PCR鉴定的影响研究  

Comparison of Five DNA Extraction Methods for Identification of Two Species Normally Caused Nosocomial Infection by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction

作　　者：魏华[1] 孙立国[1] 朱晶[1] 张晓君[1] 朱晨光[1] 赵立平[1] 庞小燕[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海210009

出　　处：《中华医院感染学杂志》2007年第9期1044-1047,共4页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金资助项目(30370031);上海市科委科研基金资助项目(05DJ14009)

摘　　要：目的研究不同基因组DNA提取方法,对肠杆菌基因间保守重复序列聚合酶链反应(ERIC-PCR)鉴定两种菌株的影响。方法采用5种基因组DNA提取法,分别为化学裂解酚/氯仿法、玻珠击打法、Triton X-100直接裂解法、Triton X-100煮沸裂解法、水煮法对1株肺炎克雷伯菌和1株大肠埃希菌基因组DNA进行提取,在相同的扩增条件下对不同方法提取的DNA模板进行ERIC-PCR。结果不同菌株基因组DNA经ERIC-PCR扩增后产生不同的电泳图谱,而用不同方法提取同一菌株DNA可以得到非常相似的电泳图谱。结论Triton X-100直接裂解法、Triton X-100煮沸裂解法、水煮法可以作为操作简单的DNA提取法用于细菌基因分型。OBJECTIVE To assess the influence of different genomic DNA extraction methods for identification of two species normally caused nosocomial infections by enterobacterial repetitive intergenic consensus polymerase chain reaction （ERIC-PCR）. METHODS The genomic DNA of one strain of Klebsiella pneurnoniae and one strain of Escherichia coli were extracted by five different methods including chemical lysis extraction, bead beating extraction, Triton X-100 extraction, Triton X-100 boiled extraction and water boiled extraction. ERIC-PCR with different DNA template was performed with the same PCR condition. RESULTS There were remarkable differences in DNA fingerprint patterns between the two different species. While, DNA fingerprint patterns of the genomie DNA from the same species extracted by different methods were very similar. CONCLUSIONS Triton X-100 extraction, Triton X-100 boiled extraction and water boiled extraction methods can be used as the simple DNA extraction methods to identify the genomic type of bacteria.

关 键 词：肺炎克雷伯菌 大肠埃希菌 DNA提取 ERIC-PCR 

分 类 号：R378[医药卫生—病原生物学]