title: Development of Multilocus Sequence Typing (MLST) for Mycoplasma synoviae.
abstract: Mycoplasma synoviae (MS) is a poultry pathogen that has had an increasing incidence and economic impact over the past few years. Strain identification is necessary for outbreak investigation, infection source identification, and facilitating prevention and control as well as eradication efforts. Currently, a segment of the variable lipoprotein hemagglutinin A (vlhA) gene (420 bp) is the only target that is used for MS strain identification. A major limitation of this assay is that colonality of typed samples can only be inferred if their vlhA sequences are identical; however, if their sequences are different, the degree of relatedness is uncertain. In this study we propose a multilocus sequence typing (MLST) assay to further refine MS strain identification. After initial screening of 24 housekeeping genes as potential targets, seven genes were selected for the MLST assay. An internal segment (450-711 bp) from each of the seven genes was successfully amplified and sequenced from 58 different MS strains and field isolates (n = 30) or positive clinical samples (n = 28). The collective sequence of all seven gene segments (3960 bp total) was used for MS sequence typing. The 58 tested MS samples were typed into 30 different sequence types using the MLST assay and, coincidentally, all the samples were typed into 30 sequence types using the vlhA assay. However, the phylogenetic tree generated using the MLST data was more congruent to the epidemiologic information than was the tree generated by the vlhA assay. We suggest that the newly developed MLST assay and the vlhA assay could be used in tandem for MS typing. The MLST assay will be a valuable and more reliable tool for MS sequence typing, providing better understanding of the epidemiology of MS infection. This in turn will aid disease prevention, control, and eradication efforts.
authors.affiliation: A Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Gazzar
authors.affiliation: A Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
authors.forename: Mostafa
authors.initials: M
authors.lastname: Ghanem
authors.affiliation: A Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
authors.forename: Kristina
authors.initials: K
authors.lastname: McDonald
authors.affiliation: B Poultry Diagnostic and Research Center, 953 College Station Road, The University of Georgia, Athens, GA 30602.
authors.forename: Naola
authors.initials: N
authors.lastname: Ferguson-Noel
authors.affiliation: A Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
authors.forename: Ziv
authors.initials: Z
authors.lastname: Raviv
authors.affiliation: A Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Slemons
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002645:Chickens; D005838:Genotype; D058885:Multilocus Sequence Typing / Q000379:methods*; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary*; D045805:Mycoplasma synoviae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D010802:Phylogeny; D011201:Poultry Diseases / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins
keywords: MLST; Mycoplasma synoviae; genotyping; multilocus sequence typing; sequence typing; vlhA gene