Strandén A, Frei R, Widmer A F
Division of Hospital Epidemiology, University Hospital Basel, CH-3031 Basel, Switzerland.
J Clin Microbiol. 2003 Jul;41(7):3181-6. doi: 10.1128/JCM.41.7.3181-3186.2003.
Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular typing of methicillin-resistant Staphylococcus aureus (MRSA). However, the method is time-consuming and expensive, and its discriminatory power may not be necessary in outbreak situations. We used a rapid multiplex PCR-based method with published primers and compared the results with those obtained by PFGE. A total of 75 clinical isolates were typed: 59 strains originated from our prospectively collected clinical strains and were epidemiologically unrelated; 16 strains came from an outbreak that was epidemiologically well defined in time and space. A primer mix of the spa gene, the coa gene, and the hypervariable region adjacent to mecA gene was used for multiplex PCR. Both PFGE and PCR clustered the 75 strains into 41 different genotypes. Concordance of the results was 100% for strains originating from the outbreak. Overall, both methods produced concordant results in 72% of cases. A total of 16% were clustered together by PFGE, but not by PCR and 12% were clustered together by PCR but not by PFGE, respectively. The turnaround time was only 8 h for PCR but 5 days for PFGE. This PCR-based method is excellent for rapid and inexpensive typing of MRSA in an outbreak setting, but the discriminatory power and reproducibility are still insufficient to replace PFGE in longitudinal studies in the endemic setting.
脉冲场凝胶电泳(PFGE)被认为是耐甲氧西林金黄色葡萄球菌(MRSA)分子分型的“金标准”。然而,该方法耗时且昂贵,在暴发情况下其鉴别能力可能并非必要。我们使用了一种基于已发表引物的快速多重PCR方法,并将结果与PFGE获得的结果进行比较。共对75株临床分离株进行分型:59株来自我们前瞻性收集的临床菌株,且在流行病学上无关联;16株来自一次在时间和空间上流行病学定义明确的暴发。使用spa基因、coa基因以及与mecA基因相邻的高变区的引物混合物进行多重PCR。PFGE和PCR均将75株菌株聚类为41种不同的基因型。来自暴发的菌株结果一致性为100%。总体而言,两种方法在72%的病例中产生了一致的结果。分别有16%的菌株通过PFGE聚类在一起,但未通过PCR聚类;12%的菌株通过PCR聚类在一起,但未通过PFGE聚类。PCR的周转时间仅为8小时,而PFGE为5天。这种基于PCR的方法在暴发情况下对MRSA进行快速且廉价的分型非常出色,但在地方性环境的纵向研究中,其鉴别能力和可重复性仍不足以取代PFGE。