Simultaneous identification of Mycoplasma gallisepticum and Mycoplasma synoviae by duplex PCR assay

JOINT EVENT ON 15^th International PHARMACEUTICAL MICROBIOLOGY AND BIOTECHNOLOGY CONFERENCE & 10^th Annual MEDICAL MICROBIOLOGY SUMMIT & EXPO

June 21-23, 2017 London, UK

Golbarg Malekhoseini

Islamic Azad University, Qom Branch, Iran

Posters & Accepted Abstracts: J Med Microb Diagn

Abstract :

Mycoplasma gallisepticum (M.G) and Mycoplasma synoviae (M.S) have recognized as common respiratory pathogens especially in chickens causing lots of economic losses in poultry industries. The aim of this study was to develop and validate duplex polymerase chain reaction (PCR) for simultaneous detection of M.G & M.S. A total of 50 samples from tracheas, lungs and air sacs were taken from commercial broiler chicken farms in Iran. The samples were cultured in PPLO broth supplemented for M.S and M.G isolation and bacteria DNA were extracted by phenol/chloroform extraction method. The conserved region of 16S rRNA gene was applied for the detection of Mycoplasma genus in 163bp fragment and M.G in 183 bp fragment and vlhA gene was also employed for detection of M.S in 350 bp fragment. Hence, duplex PCR amplified the conserved region of 16S rRNA and vlhA genes which were then applied for detection of M.G & M.S. 20 samples in Mycoplasma genus, and seven samples in M.G & M.S were positive in the single PCR whereas in three samples M.G & M.S were simultaneously positive in the duplex PCR method. The results showed that duplex PCR was successful to simultaneous identification of M.G & M.S and suggested that duplex PCR is more rapid and inexpensive method than the single PCR for detection of M.G & M.S.

Biography :

Email: g1_malekhoseini@yahoo.com