Rapid Molecular Typing of Methicillin-Resistant Staphylococcus aureus by PCR-RFLP
Rapid Molecular Typing of Methicillin-Resistant Staphylococcus aureus by PCR-RFLP
复制标题
PCR-RFLP 法对耐甲氧西林金黄色葡萄球菌进行快速分子分型
DOI：
10.1086/501903
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
V. Brade
中科院分区：
文献类型：
--
作者：
T. Wichelhaus;K. Hunfeld;B. Böddinghaus;P. Kraiczy;V. Schàfer;V. Brade
摘要目的：建立一种新的、快速、可靠的耐甲氧西林金黄色葡萄球菌（MRSA）基因型指纹分型技术，用于MRSA的常规流行病学监测。设计图：该方法是基于聚合酶链反应（PCR）限制性片段长度多态性（RFLP）的HaeII消化的同时扩增的部分蛋白A基因，凝固酶基因，和邻近mecA的高变区。共分析了46株MRSA初始分离株，包括来自5个国家的14株分离株; 6株德国流行株;来自法兰克福大都市区的16株分离株，已知脉冲场凝胶电泳（PFGE）为异质性;以及在3次流行期间获得的10株分离株，所有分离株均显示出相同的基因型。结果如下：限制性内切酶分析PCR-RFLP允许歧视的10个14个国际分离株，所有6个德国流行株，和15个16个国家分离株。这也证实了10个爆发分离株的同质性。结论：PCR-RFLP分型方法是一种新的、快速的流行病学监测方法。其令人印象深刻的特点是易于执行和解释，同时保证良好的区分能力，可重复性和可打字性。
Abstract Objective: To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistant Staphylococcus aureus (MRSA) typing in routine epidemiological surveillance. Design: The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) following HaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent to mecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype. Results: Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates. Conclusions: This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.