金黄色葡萄球菌基因组DNA提取方法的比较研究

Evaluations on DNA extraction methods of Staphylococcus aureus

目的:对金黄色葡萄球菌DNA不同提取方法进行评价。方法:采用4种不同金黄色葡萄球菌DNA提取方法,比较提取得率和质量,对提取的DNA进行PCR扩增并评价效果。结果:在DNA提取之前加入70%乙醇可有效处理细菌细胞,增加细胞对裂解液的敏感性,再用溶菌酶处理能很好地被消化,获得了一种快速、简洁和有效的提取方法。结论:这是一种适用于金黄色葡萄球菌和其它病原细菌的有效提取方法(方法4),可用于金黄色葡萄球菌和其它病原细菌的分子生物学操作和快速诊断技术。

Objective： To evaluate different extraction methods used for Staphylococcus aureus genome DNA.Methods：Four methods have been applied to extract the DNA from the S.aureus.The yield and quality were compared through agarose gel electrophoresis and PCR.Results：Bacterial cells were pretreated with 70% ethanol prior to enzymatic digestion with lysozyme which could make the cultures sensitive and increase the susceptibility of the cells to lysozyme-induced lysis.One rapid,simple extraction method has been selected and improved.Conclusion：Method 4 of this study is effective and generally applicable to other pathogenic bacteria.The significance of this study is to provide a basic molecular biology technique for DNA extraction from S.aureus and other gram-positive bacteria.