Development of a Multilocus Sequence Typing Scheme for Molecular Typing of Mycoplasma pneumoniae

Abstract

Mycoplasma pneumoniaeis a major human respiratory pathogen causing both upper and lower respiratory disease in humans of all ages, and it can also result in other serious extrapulmonary sequelae. A multilocus sequence typing (MLST) scheme forM. pneumoniaewas developed based on the sequences of eight housekeeping genes (ppa,pgm,gyrB,gmk,glyA,atpA,arcC, andadk) and applied to 55M. pneumoniaeclinical isolates and the two type strains M129 and FH. A total of 12 sequence types (STs) resulted for 57M. pneumoniaeisolates tested, with a discriminatory index of 0.21 STs per isolate. The MLST loci used in this scheme were shown to be stable in 10 strains following 10 sequential subculture passages. Phylogenetic analysis of concatenated sequences of the eight loci indicated two distinct genetic clusters that were directly linked to multilocus variable-number tandem repeat analysis (MLVA) type. Genetic MLST clustering was confirmed by genomic sequence analysis, indicating that the MLST scheme developed in this study is representative of the genome. Furthermore, this MLST scheme was shown to be more discriminatory than both MLVA and P1 typing for theM. pneumoniaeisolates examined, providing a method for further and more detailed analysis of observed epidemic peaks ofM. pneumoniaeinfection. This scheme is supported by a public Web-based database (http://pubmlst.org/mpneumoniae).