Development of a quantitative real-time PCR assay for the detection and comparison of avian Mycoplasma synoviae in chickens from MS vaccine efficacy test
SX Huang, KY Huang, HC Lin, JP Wang, ZW Chen, JH Lin, HY Chou
Mycoplasma synoviae causes exudates in the joints (arthritis), tendons and sternal bursae of infected birds. In this study, we develop a real-time polymerase chain reaction (qPCR) with an internal positive control to evaluate the protective efficacy of a M. synoviae vaccine developed in our lab. For quantitation of the copy number of M. synoviae, we first established a standard curve with 10-fold serial diluted plasmids containing the M. synoviae specific gene. We found that the sensitivity of the real-time PCR is 2500 copies of M. synoviae per reaction. The specificity of the real-time PCR assay was further verified by examining genomic DNA from different Mycoplasma and bacteria. Finally, the anti-M. synoviae antibody titer from the joint and tracheal specimens of SPF chicken received in M. synoviae vaccine and M. synoviae challenge were determined. The real-time PCR developed in the present study is highly rapid and easy to perform, and thus it might have the potential to become an alternative method for the detection and comparison of M. synoviae titer from clinical samples.