title: Molecular Epidemiology of Mycoplasma pneumoniae: Genotyping Using Single Nucleotide Polymorphisms and SNaPshot Technology.
abstract: Molecular typing of Mycoplasma pneumoniae is an important tool for identifying grouped cases and investigating outbreaks. In the present study, we developed a new genotyping method based on single nucleotide polymorphisms (SNPs) selected from the whole-genome sequencing of eight M. pneumoniae strains, using the SNaPshot minisequencing assay. Eight SNPs, localized in housekeeping genes, predicted lipoproteins, and adhesin P1 genes were selected for genotyping. These SNPs were evaluated on 140 M. pneumoniae clinical isolates previously genotyped by multilocus variable-number tandem-repeat analysis (MLVA-5) and adhesin P1 typing. This method was also adapted for direct use with clinical samples and evaluated on 51 clinical specimens. The analysis of the clinical isolates using the SNP typing method showed nine distinct SNP types with a Hunter and Gaston diversity index (HGDI) of 0.836, which is higher than the HGDI of 0.583 retrieved for the MLVA-4 typing method, where the nonstable Mpn1 marker was removed. A strong correlation with the P1 adhesin gene typing results was observed. The congruence was poor between MLVA-5 and SNP typing, indicating distinct genotyping schemes. Combining the results increased the discriminatory power. This new typing method based on SNPs and the SNaPshot technology is a method for rapid M. pneumoniae typing directly from clinical specimens, which does not require any sequencing step. This method is based on stable markers and provides information distinct from but complementary to MLVA typing. The combined use of SNPs and MLVA typing provides powerful discrimination of strains.
authors.affiliation: University of Bordeaux, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France INRA, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France.
authors.forename: A
authors.initials: A
authors.lastname: Touati
authors.affiliation: Institute of Genetics and Microbiology, University of Paris-Sud, Orsay, France CNRS, Orsay, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Blouin
authors.affiliation: University of Bordeaux, UMR 1332 Biologie du Fruit et Pathologie, Villenave d'Ornon, France INRA, UMR 1332, Biologie du Fruit et Pathologie, Villenave d'Ornon, France.
authors.forename: P
authors.initials: P
authors.lastname: Sirand-Pugnet
authors.affiliation: Bacteriology Department, Bordeaux University Hospital, Bordeaux, France.
authors.forename: H
authors.initials: H
authors.lastname: Renaudin
authors.affiliation: Department of Pediatrics, Niigata University Medical and Dental Hospital, Niigata, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Oishi
authors.affiliation: Institute of Genetics and Microbiology, University of Paris-Sud, Orsay, France CNRS, Orsay, France ENSTA ParisTech, University of Paris-Saclay, Palaiseau, France.
authors.forename: G
authors.initials: G
authors.lastname: Vergnaud
authors.affiliation: University of Bordeaux, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France INRA, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France Bacteriology Department, Bordeaux University Hospital, Bordeaux, France.
authors.forename: C
authors.initials: C
authors.lastname: Bébéar
authors.affiliation: University of Bordeaux, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France INRA, USC EA3671 Mycoplasmal and Chlamydial Infections in Humans, Bordeaux, France Bacteriology Department, Bordeaux University Hospital, Bordeaux, France sabine.pereyre@u-bordeaux.fr.
authors.forename: S
authors.initials: S
authors.lastname: Pereyre
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics; D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D005260:Female; D020043:Genes, Essential; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D007223:Infant; D008297:Male; D017720:Molecular Epidemiology / Q000379:methods; D058889:Molecular Typing / Q000379:methods*; D009177:Mycoplasma pneumoniae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D011019:Pneumonia, Mycoplasma / Q000453:epidemiology* / Q000382:microbiology*; D020641:Polymorphism, Single Nucleotide
chemical_list: D018829:Adhesins, Bacterial