Krawczyk Beata, Leibner Justyna, Barańska-Rybak Wioletta, Samet Alfred, Nowicki Roman, Kur Józef
Department of Microbiology, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-952 Gdańsk, Poland.
J Microbiol Methods. 2007 Nov;71(2):114-22. doi: 10.1016/j.mimet.2007.08.010. Epub 2007 Aug 31.
The present study was designed to evaluate the usefulness of two novel molecular typing methods, amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting) and the PCR melting profile (PCR MP), for Staphylococcus aureus strain differentiation. Thirty-seven S. aureus strains isolated from patients with a history of furunculosis were studied. The strains were identified by determining several phenotypic properties and were genotyped using three differentiation methods: macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE), ADSRRS-fingerprinting, and PCR MP technique. In some cases the results obtained showed that the S. aureus isolated from the nose was identical to the one from the furuncle of the same patient. The same genotype was also identified for S. aureus strains isolated from two different members of a family with a history of recurrent furunculosis, although the active lesions were present in only one of them when the investigation was done. Results from strain genotyping illustrated that the recently developed ADSRRS-fingerprinting and PCR MP techniques are useful for studies of intraspecies genetic relatedness of S. aureus strains. They are as effective in discriminating closely related strains as the PFGE method, which is currently considered to be "the gold standard" for epidemiological studies.
本研究旨在评估两种新型分子分型方法,即围绕稀有限制性位点的DNA片段扩增(ADSRRS指纹图谱法)和PCR熔解曲线分析(PCR MP),用于金黄色葡萄球菌菌株分型的实用性。对37株从有疖病病史患者中分离出的金黄色葡萄球菌菌株进行了研究。通过测定多种表型特性对菌株进行鉴定,并使用三种分型方法进行基因分型:脉冲场凝胶电泳对染色体DNA进行宏观限制性分析(REA-PFGE)、ADSRRS指纹图谱法和PCR MP技术。在某些情况下,所获得的结果表明,从鼻子分离出的金黄色葡萄球菌与同一患者疖肿中分离出的菌株相同。对于从有复发性疖病病史的一个家庭的两名不同成员中分离出的金黄色葡萄球菌菌株,也鉴定出了相同的基因型,尽管在进行调查时,活动性病变仅出现在其中一人身上。菌株基因分型结果表明,最近开发的ADSRRS指纹图谱法和PCR MP技术可用于金黄色葡萄球菌菌株种内遗传相关性研究。它们在区分密切相关菌株方面与PFGE方法一样有效,而PFGE方法目前被认为是流行病学研究的“金标准”。