Afanas'ev M V, Karakashev S V, Il'ina E N, Salem Al-Salami A M, Sidorenko S V, Govorun V M
Mol Gen Mikrobiol Virusol. 2010(2):20-4.
OBJECTIVES; Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen of nosocomial infection. The goal of this work was to evaluate the clonality of hospital-acquired MRSA (HA-MRSA) circulating in Russian Federation and to compare different multiplex PCR techniques with SNP-based approach for MRSA typing.
Epidemiologically unrelated MRSA isolates (n = 62) from Moscow hospitals were selected for typing. Genomic DNA from clinical isolates was purified using the DNA express kit (Lytech Ltd, Russia). Staphylococcus chromosomal cassette mec (SCCmec) typing was performed by PCR using the previously described methods. Seven loci from five housekeeping genes (arcC162, arcC210, aroE132, gmk123, tpi241, tpi243 and yqiL333) were used for SNP-typing. Detection of particular nucleotides in selected loci was carried out in the thermocyclic primer extension reaction, followed by mass spectrometry of the products. Standard MLST procedure was performed as reference method.
The majority of the MRSA isolates (93.6%) belong to world-wide disseminated clonal complex (CC) 8. Three isolates (4.8%) belong to CC 1. All ST 239 isolates were found to carry SCCmec type III; ST 8 isolates, SCCmec type IV.
Among Russian MRSA CC 8 isolates carrying SCCmec IV type are predominant. SNP-typing is powerful toll for studies of molecular epidemiology of MRSA.
目的；耐甲氧西林金黄色葡萄球菌（MRSA）是医院感染的常见病原体。本研究的目的是评估在俄罗斯联邦传播的医院获得性MRSA（HA-MRSA）的克隆性，并比较不同的多重PCR技术与基于单核苷酸多态性（SNP）的MRSA分型方法。
选择来自莫斯科医院的62株流行病学无关的MRSA分离株进行分型。使用DNA express试剂盒（俄罗斯Lytech有限公司）从临床分离株中纯化基因组DNA。采用先前描述的方法通过PCR进行葡萄球菌染色体盒式mec（SCCmec）分型。来自五个管家基因（arcC162、arcC210、aroE132、gmk123、tpi241、tpi243和yqiL333）的七个位点用于SNP分型。在热循环引物延伸反应中检测选定位点的特定核苷酸，然后对产物进行质谱分析。标准的多位点序列分型（MLST）程序作为参考方法。
大多数MRSA分离株（占93.6%）属于全球传播的克隆复合体（CC）8。三株分离株（占4.8%）属于CC 1。所有ST239分离株均携带III型SCCmec；ST8分离株携带IV型SCCmec。
在俄罗斯的MRSA中，携带IV型SCCmec的CC 8分离株占主导地位。SNP分型是研究MRSA分子流行病学强有力的工具。
