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Simultaneous identification of Mycoplasma gallisepticum and Mycoplasma synoviae by duplex PCR assay
7th World Congress on Microbiology
November 28-29, 2016 Valencia, Spain
Golbarg Malekhoseini
Islamic Azad University, Iran
Posters & Accepted Abstracts: J Bacteriol Parasitol
Abstract:
Mycoplasma gallisepticum and Mycoplasma synoviae have recognized as common respiratory pathogens especially in chickens causing lots of economical losses in poultry industries. The aim of this study was to develop and validate duplex Polymerase Chain Reaction (PCR) for simultaneous detection of M. gallisepticum and M. synoviae. 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. gallisepticum and M. synoviae 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 163 bp fragment and M. gallisepticum in 183 bp fragment and vlhA gene was also employed for detection of M. synoviae 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. gallisepticum and M. synoviae. 20 samples in Mycoplasma genus and 7 samples in M. gallisepticum and M. synoviae were positive in the single PCR whereas in 3 samples M. gallisepticum and M. synoviae were simultaneously positive in the duplex PCR method. The results showed that duplex PCR was successful to simultaneous identification of M. gallisepticum and M. synoviae and suggested that duplex PCR is more rapid and inexpensive method than the single PCR for detection of M. gallisepticum and M. synoviae.
Biography :
Golbarg Malekhoseini has completed her MSc in 2011 from University of Qom, Pakistan. Currently, she is an Assistant Professor at Islamic Azad University of Arak and also working as a Manager of Quality Control in Ice Factory of Arak.
Email: g1_malekhoseini@yahoo.com