Deplano A, Vaneechoutte M, Verschraegen G, Struelens M J
Department of Microbiology, Hôpital Erasme, School of Public Health, Université Libre de Bruxelles, Brussels, Belgium.
J Clin Microbiol. 1997 Oct;35(10):2580-7. doi: 10.1128/jcm.35.10.2580-2587.1997.
IS256 elements are present in multiple copies in the staphylococcal genome, either flanking the transposon Tn4001 or independent of it. PCR-based analysis of inter-IS256 spacer polymorphisms was developed for typing of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis strains. Using SmaI macrorestriction analysis resolved by pulsed-field gel electrophoresis (PFGE) as the reference method for MRSA typing, excellent reproducibility (100%), discriminatory power (97%), and in vivo stability were observed. Good concordance of the results with those of other molecular typing methods was found for two MRSA collections. Inter-IS256 PCR analysis of a U.S. collection of MRSA strains (n = 36), previously characterized by 15 typing methods, showed more limited discrimination. Agreement was 78% with PFGE analysis and 83% with ribotyping (HindIII). Analysis of a second set of Belgian MRSA strains (n = 17), categorized into two widespread epidemic clones by PFGE analysis, showed 65% agreement. For typing of S. epidermidis strains (n = 26), inter-IS256 PCR showed complete typeability (100%) and good discriminatory power (85%). Inter-IS256 PCR analysis is proposed as an efficient molecular typing assay for epidemiological studies of MRSA or S. epidermidis isolates.
IS256元件以多拷贝形式存在于葡萄球菌基因组中,要么位于转座子Tn4001两侧,要么与之无关。基于聚合酶链反应(PCR)的IS256间隔区多态性分析被开发用于耐甲氧西林金黄色葡萄球菌(MRSA)和表皮葡萄球菌菌株的分型。使用通过脉冲场凝胶电泳(PFGE)解析的SmaI宏限制性分析作为MRSA分型的参考方法,观察到了极好的重现性(100%)、鉴别力(97%)和体内稳定性。对于两个MRSA菌株集合,发现结果与其他分子分型方法的结果具有良好的一致性。对一组先前用15种分型方法进行表征的美国MRSA菌株(n = 36)进行的IS256 PCR分析显示鉴别力更有限。与PFGE分析的一致性为78%,与核糖体分型(HindIII)的一致性为83%。对第二组比利时MRSA菌株(n = 17)进行分析,通过PFGE分析将其分为两个广泛传播的流行克隆,一致性为65%。对于表皮葡萄球菌菌株(n = 26)的分型,IS256 PCR显示完全可分型性(100%)和良好的鉴别力(85%)。IS256 PCR分析被提议作为一种用于MRSA或表皮葡萄球菌分离株流行病学研究的有效分子分型检测方法。