Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus
Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus
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DOI：
10.1128/aac.46.7.2155-2161.2002
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发表时间：
2002-07-01
影响因子：
4.9
通讯作者：
de Lencastre, H
中科院分区：
文献类型：
--
作者：
Oliveira, DC;de Lencastre, H
甲氧西林耐药金黄色葡萄球菌（MRSA）的全面表征不仅需要定义细菌的遗传背景，而且还需要定义这些细菌携带的复杂和异源mec元件的结构，该元件与耐药决定簇mecA相关。我们报告的开发，验证和应用的多重PCR策略，允许快速推定表征的mec元素类型的基础上的结构特征，被证明是典型的mec元素所携带的几个MRSA克隆。该策略通过使用大流行性MRSA克隆的代表性集合进行验证，其中相关mec元件的完整结构先前通过杂交和PCR筛选以及DNA测序确定。该方法与多位点序列分型和其他分型方法一起进行了测试，用于表征西班牙巴塞罗那医院爆发期间回收的18株代表MRSA克隆的分离株。多重PCR被证明是快速的，稳健的，并能够在一个单一的测定中确定这些菌株中的mec元件的五种结构类型，三个主要和两个次要的变体，其中每一个已经被发现在MRSA表征较早。这种技术应该是一个有用的除了armamentarium的分子分型工具的MRSA克隆类型的表征和快速初步鉴定的mec元素的结构变体。
Full characterization of methicillin-resistant Staphylococcus aureus (MRSA) requires definition of not only the bacterial genetic background but also the structure of the complex and heterologous mec element these bacteria carry, which is associated with drug resistance determinant mecA. We report the development, validation, and application of a multiplex PCR strategy that allows quick presumptive characterization of the mec element types based on the structural features that were shown to be typical of mec elements carried by several MRSA clones. The strategy was validated by using a representative collection of pandemic MRSA clones in which the full structure of the associated mec elements was previously determined by hybridization and PCR screenings and also by DNA sequencing. The method was tested together with multillocus sequence typing and other typing methods for the characterization of 18 isolates representative of the MRSA clones recovered during a hospital outbreak in Barcelona, Spain. The multiplex PCR was shown to be rapid, robust, and capable in a single assay of identifying five structural types of the mec element among these strains, three major and two minor variants, each one of which has been already been seen among MRSA characterized earlier. This technique should be a useful addition to the armamentarium of molecular typing tools for the characterization of MRSA clonal types and for the rapid tentative identification of structural variants of the mec element.