耐甲氧西林金黄色葡萄球菌基因诊断及多重PCR SCCmec基因分型方法的建立

目的建立对临床分离的耐甲氧西林金黄色葡萄球菌(MRSA)基因诊断和金黄色葡萄球菌染色体mec基因盒(SCCmec)基因分型的方法。方法对临床分离的11株金黄色葡萄球菌采用双重PCR(femA和mecA)鉴定,再将鉴定为MRSA的临床菌株和3株标准株用多重PCR方法在一个反应体系(针对MRSA的8个基因)中进行SCCmec基因分型。结果临床分离株中有6株鉴定为MRSA,对其和MRSA标准株的多重PCR基因分型结果显示,两株临床株为SCCmecⅡ型,4株为Ⅲ型,标准株SA-w2为Ⅰ型,MRSA252为Ⅱ型。结论该研究可以很好地对临床MRSA进行基因诊断和分型,对MRSA的诊断和分型、耐药研究以及分子流行病学有重要意义。

Genetic diagnosis of MRSA and development of multiplex PCR for staphylococcal cassette chromosome mec genotyping

OBJECTIVE To develop assays for genetic diagnosis and staphylococcal cassette chromosome mec typing of meticillin-resistant Staphylococcus aureus(MRSA) from clinical samples.METHODS A total of 11 strains of S.aureus were isolated from clinical samples,the genetic identification of those isolates was by diplex PCR(femA+mecA),then SCCmec genotyping was performed for the identified MRSA strains and 3 control MRSA strains by multiplex PCR in one panel(detected 8 genes).RESULTS Totally 6 of 11 clinical strains were identified to be MRSA,SA-w2 was classified to be SCCmec typeⅠ,2 clinical MRSA strains and MRSA 252 were classified to be SCCmec typeⅡ,4 clinical MRSA strains were classified to be SCCmec type Ⅲ.CONCLUSION The genetic identification and genotyping of the clinical MRSA strains can be performed with this method which is very useful to the diagnosis,genotyping,and the research of drug resistant and molecular epidemiology of MRSA.