Specific detection and typing of Mycoplasma synoviae strains in poultry with PCR and DNA sequence analysis targeting the hemagglutinin encoding gene vlhA.
Mycoplasma synoviae is a major pathogen of chickens and turkeys, causing economic losses to the poultry industry worldwide. In this study, we validated and applied polymerase chain reaction (PCR) and DNA sequence analysis on the N-terminal end of the hemagglutinin encoding gene vlhA as an alternative for the detection and initial typing of field strains of M. synoviae in commercial poultry. PCR primers were tested against isolates of M. synoviae from various sources along with other avian mycoplasma and other bacterial species. The vlhA gene-targeted PCR assay was highly specific in the identification of M. synoviae, with a detection limit of 4.7 x 10(2) color changing units/ml. DNA sequence analysis of amplified products was also conducted to validate the potential for typing M. synoviae strains using the N-terminal region of the vlhA gene. To evaluate the test, we applied the PCR assay to tracheal swabs collected from chickens challenged with M. synoviae strain K1968 and compared the results to the serologic detection. The PCR assay was also evaluated directly on tracheal samples collected from commercial layers. Overall, this vlhA gene-targeted PCR is a useful tool for detection and initial typing of M. synoviae and can be applied in the preliminary identification of M. synoviae isolates directly from clinical samples.
Hong
Yang
Y
Department of Avian Medicine, College of Veterinary Medicine, The University of Georgia, Athens, GA 30602-4875, USA.
García
Maricarmen
M
Leiting
Victoria
V
Bencina
Dulan
D
Dufour-Zavala
Louise
L
Zavala
Guillermo
G
Kleven
Stanley H
SH
D000372:Agglutination Tests / Q000662:veterinary; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002645:Chickens / Q000235:genetics / Q000382:microbiology*; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D006388:Hemagglutinins / Q000235:genetics*; D037102:Lectins; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000235:genetics / Q000662:veterinary*; D045805:Mycoplasma synoviae / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000235:genetics / Q000276:immunology / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014422:Turkeys / Q000235:genetics / Q000382:microbiology*; D014481:United States
D001426:Bacterial Proteins; D017931:DNA Primers; D006388:Hemagglutinins; D037102:Lectins; D012336:RNA, Ribosomal, 16S; C406455:VlhA protein, Mycoplasma synoviae