Department of Bacteriology, Capital Institute of Pediatrics, Beijing, China.
Department of Bacteriology, Capital Institute of Pediatrics, Beijing, China
J Clin Microbiol. 2014 Dec;52(12):4168-71. doi: 10.1128/JCM.02178-14. Epub 2014 Sep 17.
In this study, we developed a single-tube multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) assay to type Mycoplasma pneumoniae directly from respiratory samples collected from children with respiratory infections. The multiplex PCR included four fluorescently primed VNTRs (Mpn13, Mpn14, Mpn15, and Mpn16) and was carried out in a single tube. A total of 137 M. pneumoniae-positive specimens, collected in 2013 from Beijing, China, were divided among four types (M4-5-7-2, M4-5-6-2, M3-5-6-2, and M5-5-7-2) using the amended MLVA system. The most prevalent genotype was M4-5-7-2. No correlation was found between macrolide resistance in the M. pneumoniae samples and the MLVA types. To our knowledge, this is the first study to type and analyze M. pneumoniae clinical specimens using multiplex PCR-capillary electrophoresis in a single tube. This novel low-cost method can be used to rapidly type M. pneumoniae clinical specimens directly and shows great potential for monitoring outbreaks of M. pneumoniae.
在这项研究中，我们开发了一种单管多位点可变数串联重复（VNTR）分析（MLVA）检测方法，可直接对呼吸道感染患儿的呼吸道样本进行肺炎支原体分型。该多重 PCR 包括四个荧光引物 VNTR（Mpn13、Mpn14、Mpn15 和 Mpn16），并在单个管中进行。2013 年从中国北京采集的 137 份 M. pneumoniae 阳性标本，使用改良的 MLVA 系统分为 4 种类型（M4-5-7-2、M4-5-6-2、M3-5-6-2 和 M5-5-7-2）。最常见的基因型是 M4-5-7-2。未发现 M. pneumoniae 样本中的大环内酯类耐药与 MLVA 类型之间存在相关性。据我们所知，这是首次在单管中使用多重 PCR-毛细管电泳对 M. pneumoniae 临床标本进行分型和分析。这种新型的低成本方法可用于快速直接对 M. pneumoniae 临床标本进行分型，并具有监测 M. pneumoniae 爆发的巨大潜力。
