title: Use of molecular diversity of Mycoplasma gallisepticum by gene-targeted sequencing (GTS) and random amplified polymorphic DNA (RAPD) analysis for epidemiological studies.
abstract: A total of 67 Mycoplasma gallisepticum field isolates from the USA, Israel and Australia, and 10 reference strains, were characterized by gene-targeted sequencing (GTS) analysis of portions of the putative cytadhesin pvpA gene, the cytadhesin gapA gene, the cytadhesin mgc2 gene, and an uncharacterized hypothetical surface lipoprotein-encoding gene designated genome coding DNA sequence (CDS) MGA_0319. The regions of the surface-protein-encoding genes targeted in this analysis were found to be stable within a strain, after sequencing different in vitro passages of M. gallisepticum reference strains. Gene sequences were first analysed on the basis of gene size polymorphism. The pvpA and mgc2 genes are characterized by the presence of different nucleotide insertions/deletions. However, differentiation of isolates based solely on pvpA/mgc2 PCR size polymorphism was not found to be a reliable method to differentiate among M. gallisepticum isolates. On the other hand, GTS analysis based on the nucleotide sequence identities of individual and multiple genes correlated with epidemiologically linked isolates and with random amplified polymorphic DNA (RAPD) analysis. GTS analysis of individual genes, gapA, MGA_0319, mgc2 and pvpA, identified 17, 16, 20 and 22 sequence types, respectively. GTS analysis using multiple gene sequences mgc2/pvpa and gapA/MGA_0319/mgc2/pvpA identified 38 and 40 sequence types, respectively. GTS of multiple surface-protein-encoding genes showed better discriminatory power than RAPD analysis, which identified 36 pattern types from the same panel of M. gallisepticum strains. These results are believed to provide the first evidence that typing of M. gallisepticum isolates by GTS analysis of surface-protein genes is a sensitive and reproducible typing method and will allow rapid global comparisons between laboratories.
authors.affiliation: Department of Avian Medicine, Poultry Diagnostic and Research Center, The University of Georgia, Athens, GA 30602-4875, USA.
authors.forename: Naola M
authors.initials: NM
authors.lastname: Ferguson
authors.affiliation: Simbios Biotecnologia, Canoas, Rio Grande do Sul, Brazil.|Universidade Luterana do Brasil, Canoas, Rio Grande do Sul, Brazil.
authors.forename: Diego
authors.initials: D
authors.lastname: Hepp
authors.affiliation: Department of Avian Medicine, Poultry Diagnostic and Research Center, The University of Georgia, Athens, GA 30602-4875, USA.
authors.forename: Shulei
authors.initials: S
authors.lastname: Sun
authors.affiliation: Simbios Biotecnologia, Canoas, Rio Grande do Sul, Brazil.|Universidade Luterana do Brasil, Canoas, Rio Grande do Sul, Brazil.
authors.forename: Nilo
authors.initials: N
authors.lastname: Ikuta
authors.affiliation: Division of Avian and Aquatic Diseases, Kimron Veterinary Institute, Beit Dagan 50250, Israel.
authors.forename: Sharon
authors.initials: S
authors.lastname: Levisohn
authors.affiliation: Department of Avian Medicine, Poultry Diagnostic and Research Center, The University of Georgia, Athens, GA 30602-4875, USA.
authors.forename: Stanley H
authors.initials: SH
authors.lastname: Kleven
authors.affiliation: Department of Avian Medicine, Poultry Diagnostic and Research Center, The University of Georgia, Athens, GA 30602-4875, USA.
authors.forename: Maricarmen
authors.initials: M
authors.lastname: García
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics; D001315:Australia; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005798:Genes, Bacterial; D014644:Genetic Variation; D005838:Genotype; D007557:Israel; D008074:Lipoproteins / Q000235:genetics; D017720:Molecular Epidemiology / Q000379:methods*; D008969:Molecular Sequence Data; D045686:Mycoplasma gallisepticum / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA; D014481:United States
chemical_list: D018829:Adhesins, Bacterial; D004269:DNA, Bacterial; C117938:GapA cytadhesin; D008074:Lipoproteins; C410583:pvpA protein, Mycoplasma gallisepticum