Molecular characterization of genomic DNA in mycoplasma pneumoniae strains isolated from serious mycoplasma pneumonia cases in 2016, Yunnan, China

Mycoplasma pneumoniae (MP) is particularly prevalent in low-immunity school-age children. Few data have been reported on MP prevalence in Yunnan, China. This study was designed to investigate the prevalence and characterize genomic DNA of MP in a small outbreak in 2016, Southwest China. RepMP4 and RepMP2/3 genes of MP positive samples were amplified for molecular typing through sequence alignment and PCR-RFLP assay. Phylogenetic trees were constructed by MEGA5.0. The results showed that two distinct P1 types (type I and type II) were prevalent in this MP outbreak. Type I was the most prevalent type, and clustered in the same evolutionary branch of C26 (China, 2012). Only 1 MP isolate belonged to type II, and clustered in the branch of KCH405 (Japan, 2016). Fifty-nine nucleotide mutations were observed in P1 genes of type I isolates (51 in RepMP4, 8 in RepMP2/3). Ninety-five nucleotide mutations were observed in P1 genes of the type II isolates (33 in RepMP4, 62 in RepMP2/3). It is noteworthy that 31 mutation sites were clustered in an 84-bp fragment in the RepMP4 gene of type II isolates. One new fragment that appeared in two of the type I strains was not found in NCBI. Nucleotide diversity analyze results showed that RepMP4 was more likely to be genetically diverse than RepMP2/3. Two-tailed Z-test result of RepMP4 suggested positive selection between 6 P1 type I isolates and M29 (China, 2005). According to secondary structure prediction, 36 new possible protein binding sites were found and another 9 sites were lost, 2 helices were missed and 1 new helix appeared in type I isolates. As for type II isolates, 16 protein binding regions were gained and 31 were lost. This study may help to understand the intrinsic geographical relatedness and contributes further to the research of MP.