Genotyping of Mycoplasma pneumoniae Clinical Isolates Reveals Eight P1 Subtypes within Two Genomic Groups

Abstract

Three methods for genotyping of Mycoplasma pneumoniaeclinical isolates were applied to 2 reference strains and 21 clinical isolates. By a modified restriction fragment length polymorphism (RFLP) analysis of PCR products of the M. pneumoniae cytadhesin P1 gene, 5 subtypes were discriminated among 13 P1 type 1 strains and 3 subtypes were discriminated among 8 P1 type 2 strains. Sequence analysis of the 16S-23S rRNA gene spacer region and part of the 23S rRNA gene revealed one nucleotide difference in the intergenic spacer region in 3 of the 21 isolates. In the 23S rRNA gene sequence of the 8 P1 type 2 strains an extra adenosine was present, but it was absent from the 13 P1 type 1 strains. On the basis of M. pneumoniae genome sequence data, primers were designed to amplify large interrepeat fragments by long PCR, and these fragments were subsequently analyzed by RFLP analysis. Only two types, long PCR types 1 and 2, could be discriminated among the M. pneumoniaeisolates. All P1 type 1 strains were assigned to long PCR type 1, and all P1 type 2 strains were assigned to long PCR type 2. These data obtained by three independent typing methods thus confirm the existence of two distinct M. pneumoniae genomic groups but expand the possibility of strain typing on the basis of variations within their P1 genes.

Mycoplasma pneumoniae from the Respiratory Tract and Beyond

“…A NASBA assay was designed to detect differences between the subtypes using two type-specific probes (445). Sequencing of the 750-bp fragment of the entire 16S-23S rRNA gene spacer region and partial 23S rRNA gene revealed an extra adenosine in subtype 2 strains that was absent from the subtype 1 strains, which has also been used for distinguishing them (440). Spuesens et al used pyrosequencing to discriminate the two subtypes based on an SNP in MPN528a and another SNP in the conserved region of the P1-coding gene (431).…”

“…P1 gene sequencing, performed by various methods, was the most commonly used genotyping method until newer methods were developed (35,36,38,345,439,440). Early techniques used RFLP or pulsed-field gel electrophoresis (PFGE) (441).…”

“…All methods allow detection of the two major subtypes as well as some variants. RFLP uses several enzymes to digest the amplicons and group them based on their band patterns (440,441,443). Denaturing gradient gel electrophoresis (DGGE), which can detect single base differences, can also be employed to distinguish P1 PCR products (38).…”

Epidemiology and mechanism of drug resistance of Mycoplasma pneumoniae in Beijing, China: A multi-center study

“…This periodicity in M. pneumoniae infection may be related to changes in the sequence of P1 adhesin, which is the main method of M. pneumoniae typing. According to the differences in the sequence of P1 adhesin gene MPN141, M. pneumoniae is divided into P1 type I and P1 type II with several subtypes [16,17]. The prevalence of M. pneumoniae subtypes differs between countries and years, and the dominance of one subtype is followed by the dominance of another M. pneumoniae subtype [18].…”

Positively interacting strains that circulate in a network structured population induce cycling epidemics of Mycoplasma Pneumoniae

“…Analysis of laboratory reports of MP infections in England and Wales from 1975 to 2009 (Nguipdop-Djomo et al ., 2013) has indicated that these epidemics last on average 18 months occurring at approximately four yearly intervals. M. pneumoniae is a polymorphic pathogen (Dorigo-Zetsma et al ., 2000; Pereyre et al ., 2012): for example, Chalker et al . (2011) identified eleven strain types circulating in England and Wales during October 2010 to January 2011.…”