title: Identification of Strain-Specific Sequences That Distinguish a Mycoplasma gallisepticum Vaccine Strain from Field Isolates.
abstract: Despite attempts to control avian mycoplasmosis through management, vaccination, and surveillance, Mycoplasma gallisepticum continues to cause significant morbidity, mortality, and economic losses in poultry production. Live attenuated vaccines are commonly used in the poultry industry to control avian mycoplasmosis; unfortunately, some vaccines may revert to virulence and vaccine strains are generally difficult to distinguish from natural field isolates. In order to identify genome differences among vaccine revertants, vaccine strains, and field isolates, whole-genome sequencing of the M. gallisepticum vaccine strain ts-11 and several "ts-11-like" strains isolated from commercial flocks was performed using Illumina and 454 pyrosequencing and the sequenced genomes compared to the M. gallisepticum Rlowreference genome. The collective contigs for each strain were annotated using the fully annotated Mycoplasma reference genome. The analysis revealed genetic differences among vlhA alleles, as well as among genes annotated as coding for a cell wall surface anchor protein (mg0377) and a hypothetical protein gene, mg0359, unique to M. gallisepticum ts-11 vaccine strain. PCR protocols were designed to target 5 sequences unique to the M. gallisepticum ts-11 strain: vlhA3.04a, vlhA3.04b, vlhA3.05, mg0377, and mg0359 All ts-11 isolates were positive for the five gene alleles tested by PCR; however, 5 to 36% of field isolates were also positive for at least one of the alleles tested. A combination of PCR tests for vlhA3.04a, vlhA3.05, and mg0359 was able to distinguish the M. gallisepticum ts-11 vaccine strain from field isolates. This method will further supplement current approaches to quickly distinguish M. gallisepticum vaccine strains from field isolates.
authors.affiliation: Poultry Diagnostic and Research Center, University of Georgia, Athens, Georgia, USA.
authors.forename: Camir
authors.initials: C
authors.lastname: Ricketts
authors.affiliation: Department of Veterinary Medicine, Universidade Federal do Parana, Curitiba, PR, Brazil.|CAPES Foundation, Ministry of Education of Brazil, Brasília, DF, Brazil.
authors.forename: Larissa
authors.initials: L
authors.lastname: Pickler
authors.affiliation: Poultry Diagnostic and Research Center, University of Georgia, Athens, Georgia, USA.
authors.forename: John
authors.initials: J
authors.lastname: Maurer
authors.affiliation: Institute of Bioinformatics, University of Georgia, Athens, Georgia, USA.
authors.forename: Saravanaraj
authors.initials: S
authors.lastname: Ayyampalayam
authors.affiliation: Poultry Diagnostic and Research Center, University of Georgia, Athens, Georgia, USA.
authors.forename: Maricarmen
authors.initials: M
authors.lastname: García
authors.affiliation: Poultry Diagnostic and Research Center, University of Georgia, Athens, Georgia, USA naolaf@uga.edu.
authors.forename: Naola M
authors.initials: NM
authors.lastname: Ferguson-Noel
mesh_terms: D000818:Animals; D001428:Bacterial Vaccines / Q000009:adverse effects / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics*; D003937:Diagnosis, Differential; D005798:Genes, Bacterial; D014644:Genetic Variation; D016680:Genome, Bacterial; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011200:Poultry; D011201:Poultry Diseases / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D014613:Vaccines, Attenuated / Q000009:adverse effects / Q000235:genetics; D014730:Veterinary Medicine / Q000379:methods
chemical_list: D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D014613:Vaccines, Attenuated
keywords: Mycoplasma gallisepticum; PCR; genomes; strain typing; ts-11; ts-11-like