Genotyping Mycoplasma synoviae: Development of a multi-locus variable number of tandem-repeats analysis and comparison with current molecular typing methods

• Published in: Veterinary microbiology Vol. 226; pp. 41 - 49
• Main Authors: Kreizinger, Zsuzsa, Sulyok, Kinga M., Bekő, Katinka, Kovács, Áron B., Grózner, Dénes, Felde, Orsolya, Marton, Szilvia, Bányai, Krisztián, Catania, Salvatore, Benčina, Dušan, Gyuranecz, Miklós
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-11-2018; Elsevier BV
• Subjects: Animals; Assaying; Bacteria; Bacterial Proteins - genetics; Chickens; Chicks; Comparative analysis; Complementation; Discrimination; Eggs; Genetic diversity; Genetic Variation; Genomes; Genotype; Genotype & phenotype; Genotypes; Genotyping; Genotyping Techniques; Hatching; International trade; Juveniles; Lectins - genetics; MLST; MLVA; Molecular Typing - methods; Multilocus Sequence Typing - methods; Mycoplasma Infections - microbiology; Mycoplasma Infections - veterinary; Mycoplasma synoviae; Mycoplasma synoviae - classification; Mycoplasma synoviae - genetics; Nucleotide sequence; Phylogeny; Sequence Analysis, DNA; Strains (organisms); Tandem Repeat Sequences - genetics; Vaccine strain; Vaccines; Vaccines, Attenuated - genetics; vlhA; vlhA; Discrimination; MLST; Vaccine strain; Mycoplasma synoviae; MLVA
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/j.vetmic.2018.10.012

•MLVA was developed for the genetic characterisation of Mycoplasma synoviae.•The MLVA assay discriminated 35 genotypes among 86 strains using seven VNTR loci.•Results showed high congruency compared with MLST and vlhA gene sequencing data.•MLVA is a highly discriminative, rapid and cost-effective alternative for MLST.•MLVA may help solving ambiguous cases in live vaccine strain differentiating assays.
Control of one of the most important avian mycoplasmas, Mycoplasma synoviae, and tracing the spread of the infection can be challenging as the pathogen is transmissible by both horizontal and vertical routes, and it can be disseminated through long distances via the hatching eggs, day-old chicks or pullets during intensive international trade. Genetic information provided by molecular typing methods support control programmes and epizootiologic studies. The aims of the present study were to develop a multi-locus variable number of tandem-repeats analysis (MLVA) method for the typing of M. synoviae isolates and to evaluate the currently used molecular typing methods which are applicable directly on clinical samples.
Tandem repeat (TR) regions were selected from the whole genome sequence of the M. synoviae type strain (WVU1853) to characterise the genetic diversity of 86 M. synoviae strains originating from 15 countries. The strains were also submitted to multi-locus sequence typing (MLST) assays, vlhA gene sequence analysis and to assays designed to differentiate live vaccine strains from field strains.
The developed MLVA involves the examination of seven TR regions and provides similar genetic resolution as the tested MLST assays by identifying 35 genotypes among the tested strains. Differentiation of the live vaccine strains from field strains was also successful with the developed assay.
The provided MLVA method proved to be a highly discriminative, rapid and cost-effective alternative typing technique for the genetic characterisation of M. synoviae and it is also suitable for the complementation of live vaccine strain differentiating assays in ambiguous cases.