Del Vecchio V G, Petroziello J M, Gress M J, McCleskey F K, Melcher G P, Crouch H K, Lupski J R
Molecular Biology Institute, University of Scranton, Pennsylvania 18510, USA.
J Clin Microbiol. 1995 Aug;33(8):2141-4. doi: 10.1128/jcm.33.8.2141-2144.1995.
Methicillin resistance in Staphylococcus aureus is a frequent cause of nosocomial and community-acquired infections. Accurate, rapid epidemiologic typing is crucial to the identification of the source and spread of infectious disease and could provide detailed information on the generation of methicillin-resistant S. aureus (MRSA) strains. The high degree of genetic relatedness of MRSA strains has precluded the use of more conventional methods of genetic fingerprinting. A rapid DNA fingerprinting method that exploits PCR amplification from a DNA repeat sequence in MRSA is described. The random chromosomal distribution of this repeat sequence provides an ideal target for detecting DNA fragment patterns specific to individual MRSA strains. Two PCR fingerprinting methods which use an oligonucleotide primer based on a repetitive sequence found in Mycoplasma pneumoniae are presented. The repetitive element sequence-based PCR (rep-PCR) and fluorophore-enhanced rep-PCR (FERP) can identify epidemic strains among background MRSA. The combination of oligonucleotide primers labeled with different fluorescent dyes allowed simultaneous FERP fingerprinting and mecA gene detection. Eight different fingerprint patterns were observed in MRSA strains collected from different sources. These techniques provide a rapid discriminatory means of molecular epidemiologic typing of MRSA involved in nosocomial infections.
金黄色葡萄球菌中的耐甲氧西林特性是医院获得性感染和社区获得性感染的常见原因。准确、快速的流行病学分型对于识别传染病的来源和传播至关重要，并且可以提供有关耐甲氧西林金黄色葡萄球菌（MRSA）菌株产生的详细信息。MRSA菌株的高度遗传相关性使得更传统的基因指纹识别方法无法使用。本文描述了一种利用MRSA中DNA重复序列进行PCR扩增的快速DNA指纹识别方法。该重复序列的随机染色体分布为检测特定于单个MRSA菌株的DNA片段模式提供了理想靶点。本文介绍了两种基于肺炎支原体中发现的重复序列使用寡核苷酸引物的PCR指纹识别方法。基于重复元件序列的PCR（rep-PCR）和荧光团增强rep-PCR（FERP）可以在背景MRSA中识别流行菌株。用不同荧光染料标记的寡核苷酸引物的组合允许同时进行FERP指纹识别和mecA基因检测。在从不同来源收集的MRSA菌株中观察到八种不同的指纹模式。这些技术为医院感染中涉及的MRSA分子流行病学分型提供了一种快速鉴别手段。
