Novel multiplex PCR assay for characterization and concomitant subtyping of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus
Novel multiplex PCR assay for characterization and concomitant subtyping of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus
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DOI：
10.1128/jcm.43.10.5026-5033.2005
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发表时间：
2005-10-01
影响因子：
9.4
通讯作者：
Conly, JM
中科院分区：
文献类型：
--
作者：
Zhang, KY;McClure, JA;Conly, JM
葡萄球菌盒式染色体mec（SCCmec）分型对于了解耐甲氧西林金黄色葡萄球菌（MRSA）的分子流行病学至关重要。SCCmec元件目前根据mec和ccr基因复合物的性质分为I至V型，并根据其垃圾场区域DNA片段进一步分为亚型。先前描述的传统SCCmec PCR分型方案需要多个引物组和PCR实验，而先前公开的多重PCR测定在其检测最近发现的类型和亚型如SCCmec V型和亚型IVa、B、c和d的能力方面受到限制。我们设计了一套新的SCCmec型和亚型独特的特异性引物，并开发了一种新的多重PCR检测方法，允许同时检测甲氧西林耐药性（mecA基因）（也用作内部对照），以便于检测和分类所有目前描述的SCCmec类型和亚型1、11、111、IVa、B、c、d，当与先前描述的分型方法相比时，我们的测定法在准确表征属于各种已知SCCmec类型和亚型的54种MRSA菌株方面表现出100%的灵敏度和特异性。通过对453株临床分离菌株的检测，证实了该方法的可行性和实用性。这种新的检测方法为MRSA的SCCmec分型提供了一种快速，简单和可行的方法，并可作为临床医生和流行病学家预防和控制这种微生物引起的感染的有用工具。
Staphylococcal cassette chromosome mec (SCCmec) typing is essential for understanding the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA). SCCmec elements are currently classified into types I to V based on the nature of the mec and ccr gene complexes, and are further classified into subtypes according to their junkyard region DNA segments. Previously described traditional SCCmec PCR typing schemes require multiple primer sets and PCR experiments, while a previously published multiplex PCR assay is limited in its ability to detect recently discovered types and subtypes such as SCCmec type V and subtypes IVa, b, c, and d. We designed new sets of SCCmec type- and subtype-unique and specific primers and developed a novel multiplex PCR assay allowing for concomitant detection of the methicillin resistance (mecA gene) (also serving as an internal control) to facilitate detection and classification of all currently described SCCmec types and subtypes 1, 11, 111, IVa, b, c, d, and V. Our assay demonstrated 100% sensitivity and specificity in accurately characterizing 54 MRSA strains belonging to the various known SCCmec types and subtypes, when compared with previously described typing methods. Further application of our assay in 453 randomly selected local clinical isolates confirmed its feasibility and practicality. This novel assay offers a rapid, simple, and feasible method for SCCmec typing of MRSA, and may serve as a useful tool for clinicians and epidemiologists in their efforts to prevent and control infections caused by this organism.