Department of Medical Microbiology, Maastricht University Medical Center, Maastricht, The Netherlands.
Diagn Microbiol Infect Dis. 2009 Dec;65(4):384-91. doi: 10.1016/j.diagmicrobio.2009.08.006. Epub 2009 Sep 24.
For us to assess the spread of methicillin-resistant Staphylococcus aureus (MRSA), typing of the staphylococcal cassette chromosome mec (SCCmec) is a valuable addition to existing typing methods, such as multilocus sequence typing (MLST). Traditional SCCmec typing assays, that is, that of Oliveira et al. and Ito et al., are polymerase chain reaction (PCR) based, requiring electrophoresis. We introduce a rapid, 2-well, multiplex real-time PCR assay that can be used directly on bacterial suspensions and is able to characterize SCCmec type I to V based on the detection of the ccr genes and the mec complex. The assay was evaluated on 212 clinical MRSA isolates from various countries, associated with MLST clonal complexes (CC) 1, 5, 8, 22, 30, and 45, as well as pig-associated CC398. When comparing the real-time PCR assay with traditional methods, the correct SCCmec element was identified in 209 (99%) of the 212 MRSA isolates. The new assay enables high-throughput analyses for SCCmec on large strain collections.
为了评估耐甲氧西林金黄色葡萄球菌（MRSA）的传播情况，葡萄球菌盒式染色体 mec（SCCmec）的分型是对现有分型方法（如多位点序列分型（MLST））的有益补充。传统的 SCCmec 分型检测方法，即 Oliveira 等人和 Ito 等人的方法，基于聚合酶链反应（PCR），需要电泳。我们引入了一种快速、双孔、多重实时 PCR 检测方法，可直接用于细菌悬浮液，并能够根据 ccr 基因和 mec 复合物的检测来表征 SCCmec 类型 I 至 V。该检测方法评估了来自不同国家的 212 株临床 MRSA 分离株，这些分离株与 MLST 克隆复合体（CC）1、5、8、22、30 和 45 以及与猪相关的 CC398 相关。当将实时 PCR 检测方法与传统方法进行比较时，212 株 MRSA 分离株中的 209 株（99%）正确鉴定了 SCCmec 元素。新的检测方法可用于对大量菌株进行 SCCmec 的高通量分析。
