title: Molecular typing of Japanese field isolates and live commercial vaccine strain of Mycoplasma synoviae using improved pulsed-field gel electrophoresis and vlhA gene sequencing.
abstract: In the present study, pulsed-field gel electrophoresis (PFGE) and vlhA gene sequence analysis were applied and verified for typing the Mycoplasma synoviae live vaccine MS-H strain and field isolates from diseased chickens in Japan. The previously published PFGE protocol using SmaI digestion could not allow the discrimination of two of the 11 M. synoviae field isolates from the vaccine strain and had relatively low discrimination power (D = 0.885). On the other hand, our new PFGE protocols using BlnI and BamHI digestions as well as the vlhA sequence analysis allowed the discrimination of all 11 M. synoviae field isolates from the vaccine strain. In addition, these PFGE protocols using BlnI and BamHI digestions generated unique fragment patterns in epidemiologically unrelated isolates, including those with identical SmaI-digested patterns or vlhA gene sequences (D = 0.987 and 1.000, respectively), and generated indistinguishable or closely related patterns in epidemiologically related isolates. Therefore, we believe that they would be useful tools to determine whether M. synoviae clinical isolates from diseased chickens are derived from the vaccine strain or wild-type strain and to further elucidate the epidemiology of M. synoviae infection.
authors.affiliation: Nippon Veterinary and Life Science University, 1-7-1, Kyonan-cho, Musashino, Tokyo 180-8602, Japan. k-harada@nvlu.ac.jp
authors.forename: Kazuki
authors.initials: K
authors.lastname: Harada
authors.forename: Mayumi
authors.initials: M
authors.lastname: Kijima-Tanaka
authors.forename: Mariko
authors.initials: M
authors.lastname: Uchiyama
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Koji
authors.initials: K
authors.lastname: Oishi
authors.forename: Megumi
authors.initials: M
authors.lastname: Arao
authors.forename: Toshio
authors.initials: T
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods / Q000662:veterinary; D001428:Bacterial Vaccines / Q000276:immunology*; D001483:Base Sequence; D002645:Chickens; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field / Q000662:veterinary; D007564:Japan / Q000453:epidemiology; D037102:Lectins / Q000235:genetics*; D009175:Mycoplasma Infections / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D045805:Mycoplasma synoviae / Q000302:isolation & purification*; D010802:Phylogeny; D011201:Poultry Diseases / Q000382:microbiology*; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D037102:Lectins; C406455:VlhA protein, Mycoplasma synoviae