The Mycoplasma gallisepticum 16S–23S rRNA Intergenic Spacer Region Sequence as a Novel Tool for Epizootiological Studies

• Published in: Avian diseases 2007-06, Vol.51 (2), p.555-560
• Main Authors: Raviv, Ziv, Callison, S, Ferguson-Noel, N, Laibinis, V, Wooten, R, Kleven, S. H
• Format: Article
• Language: English
• Subjects: 16S rRNA; 23S rRNA; Animals; Bird Diseases - diagnosis; Bird Diseases - microbiology; DNA, Ribosomal Spacer - genetics; epizootiology; genetic polymorphism; Genomes; Intergenic DNA; intergenic spacer region; internal transcribed spacers; isolate; loci; microbial genetics; molecular sequence data; Mollicutes; Mycoplasma; Mycoplasma gallisepticum; Mycoplasma gallisepticum - classification; Mycoplasma gallisepticum - genetics; Mycoplasma gallisepticum - isolation & purification; Mycoplasma Infections - diagnosis; Mycoplasma Infections - microbiology; Mycoplasma Infections - veterinary; mycoplasmosis; Operator regions; operon; Phylogeny; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; poultry; poultry diseases; Regular s; ribosomal RNA; RNA, Ribosomal, 16S - genetics; RNA, Ribosomal, 23S - genetics; rRNA genes; Sensitivity and Specificity; Sequencing; single-locus sequence typing; strain; Vaccination
• ISSN: 0005-2086; 1938-4351
• DOI: 10.1637/0005-2086(2007)51[555:TMGSRI]2.0.CO;2

Mycoplasma gallisepticum (MG) contains two sets of rRNA genes (5S, 16S and 23S) in its genome, but only one of the two is organized in an operon cluster and contains a unique 660-nucleotide intergenic spacer region (IGSR) between the 16S and the 23S rRNA genes. We designed a polymerase chain reaction (PCR) for the specific amplification of the complete MG IGSR segment. The MG IGSR PCR was tested on 18 avian mollicute species and was confirmed as MG specific. The reaction sensitivity was demonstrated by comparing it to the well-established MG mgc2 PCR. The MG IGSR sequence was found to be highly variable (discrimination [D] index of 0.950) among a variety of MG laboratory strains, vaccine strains, and field isolates. The sequencing of the MG IGSR appears to be a valuable single-locus sequence typing (SLST) tool for MG isolate differentiation in diagnostic cases and epizootiological studies.