A single-tube multiple-locus variable-number tandem-repeat analysis of Mycoplasma pneumoniae clinical specimens by use of multiplex PCR-capillary electrophoresis.

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.