Walker J, Borrow R, Edwards-Jones V, Oppenheim B A, Fox A J
Manchester Public Health Laboratory, Withington Hospital, West Didsbury.
Epidemiol Infect. 1998 Dec;121(3):507-14. doi: 10.1017/s0950268898001538.
In a comparative study, isolates of methicillin-resistant Staphylococcus aureus (MRSA) with known pulsed-field gel electrophoresis (PFGE) and bacteriophage type were analysed by polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP) for additional discriminatory subtyping information. PFGE was previously performed using standardized, commercially available kits and pre-programmed software. Isolates were examined for coagulase (coa) and protein A (spa) gene polymorphisms following PCR amplification of the coa hypervariable and spa repeat regions. Coa gene RFLPs produced a total of 38 distinct combined patterns after digestion with HaeIII and AluI and identified the predominant epidemic (EMRSA) types 15 and 16. A unique HaeIII restriction site was identified by RFLP and sequence analysis in the coa gene for EMRSA 15 but not EMRSA 16. The spa gene PCR yielded a total of 14 different profiles ranging from 3-18 repeats with the 2 predominant EMRSA types falling into 2 distinct groups. PCR detection of coa and spa polymorphisms offer a rapid preliminary strain identification and discriminatory subtyping information for surveillance of MRSA.
在一项比较研究中,对已知脉冲场凝胶电泳(PFGE)和噬菌体分型的耐甲氧西林金黄色葡萄球菌(MRSA)分离株进行聚合酶链反应(PCR)和限制性片段长度多态性(RFLP)分析,以获取更多鉴别性亚型信息。PFGE先前使用标准化的市售试剂盒和预编程软件进行。在对coa高变区和spa重复区进行PCR扩增后,检测分离株的凝固酶(coa)和蛋白A(spa)基因多态性。用HaeIII和AluI消化后,coa基因RFLP共产生38种不同的组合模式,并鉴定出主要的流行(EMRSA)类型15和16。通过RFLP和序列分析在coa基因中鉴定出EMRSA 15的一个独特HaeIII限制性位点,但EMRSA 16没有。spa基因PCR共产生14种不同的图谱,重复次数从3到18次不等,两种主要的EMRSA类型分为两个不同的组。PCR检测coa和spa多态性为MRSA监测提供了快速的初步菌株鉴定和鉴别性亚型信息。