Deplano A, Schuermans A, Van Eldere J, Witte W, Meugnier H, Etienne J, Grundmann H, Jonas D, Noordhoek G T, Dijkstra J, van Belkum A, van Leeuwen W, Tassios P T, Legakis N J, van der Zee A, Bergmans A, Blanc D S, Tenover F C, Cookson B C, O'Neil G, Struelens M J
Reference Laboratory for Staphylococci, Université Libre de Bruxelles, Hôpital Erasme, 1070 Brussels, Belgium.
J Clin Microbiol. 2000 Oct;38(10):3527-33. doi: 10.1128/JCM.38.10.3527-3533.2000.
Rapid and efficient epidemiologic typing systems would be useful to monitor transmission of methicillin-resistant Staphylococcus aureus (MRSA) at both local and interregional levels. To evaluate the intralaboratory performance and interlaboratory reproducibility of three recently developed repeat-element PCR (rep-PCR) methods for the typing of MRSA, 50 MRSA strains characterized by pulsed-field gel electrophoresis (PFGE) (SmaI) analysis and epidemiological data were blindly typed by inter-IS256, 16S-23S ribosomal DNA (rDNA), and MP3 PCR in 12 laboratories in eight countries using standard reagents and protocols. Performance of typing was defined by reproducibility (R), discriminatory power (D), and agreement with PFGE analysis. Interlaboratory reproducibility of pattern and type classification was assessed visually and using gel analysis software. Each typing method showed a different performance level in each center. In the center performing best with each method, inter-IS256 PCR typing achieved R = 100% and D = 100%; 16S-23S rDNA PCR, R = 100% and D = 82%; and MP3 PCR, R = 80% and D = 83%. Concordance between rep-PCR type and PFGE type ranged by center: 70 to 90% for inter-IS256 PCR, 44 to 57% for 16S-23S rDNA PCR, and 53 to 54% for MP3 PCR analysis. In conclusion, the performance of inter-IS256 PCR typing was similar to that of PFGE analysis in some but not all centers, whereas other rep-PCR protocols showed lower discrimination and intralaboratory reproducibility. None of these assays, however, was sufficiently reproducible for interlaboratory exchange of data.
快速有效的流行病学分型系统对于监测耐甲氧西林金黄色葡萄球菌（MRSA）在本地和区域间的传播将非常有用。为了评估三种最近开发的用于MRSA分型的重复元件PCR（rep-PCR）方法在实验室内的性能和实验室间的可重复性，采用标准试剂和方案，在八个国家的12个实验室中，对50株通过脉冲场凝胶电泳（PFGE）（SmaI）分析和流行病学数据进行特征鉴定的MRSA菌株，采用IS256内部、16S-23S核糖体DNA（rDNA）和MP3 PCR进行盲法分型。分型性能通过可重复性（R）、鉴别力（D）以及与PFGE分析的一致性来定义。通过视觉和使用凝胶分析软件评估模式和类型分类的实验室间可重复性。每种分型方法在每个中心都表现出不同的性能水平。在每种方法表现最佳的中心，IS256内部PCR分型的R = 100%，D = 100%；16S-23S rDNA PCR的R = 100%，D = 82%；MP3 PCR的R = 80%，D = 83%。rep-PCR类型与PFGE类型之间的一致性因中心而异：IS256内部PCR为70%至90%，16S-23S rDNA PCR为44%至57%，MP3 PCR分析为53%至54%。总之，IS256内部PCR分型在一些但并非所有中心的性能与PFGE分析相似，而其他rep-PCR方案显示出较低的鉴别力和实验室内可重复性。然而，这些检测方法中没有一种对于实验室间的数据交换具有足够的可重复性。
