Journal of Microbial & Biochemical Technology

Comparison between different molecular methods for detection of Mycoplasma gallisepticum and Mycoplasma synoviae

Joint Event on 4^th World Congress and Expo on Applied Microbiology & 2^nd International Conference on Food Microbiology

November 29-December 01, 2017 Madrid, Spain

Golbarg Malekhoseini

Islamic Azad University, Qom Branch, Iran

Posters & Accepted Abstracts: J Microb Biochem Technol

Abstract:

Avian mycoplasmosis is caused by several pathogenic mycoplasmas of which Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) have long been recognized as common respiratory pathogens especially in chickens. The aim of this study was comparison of different methods such as Culture, Serological Methods, PCR, Duplex PCR, RAPD PCR and REAL TIME-PCR for detection of Mycoplasma Gallisepticum and Mycoplasma Synoviae. The result of this study was shown that duplex PCR has more specificity than other methods and it is more rapid and inexpensive method than other methods for detection of MG and MS so duplex PCR can be an alternative method for detection MG and MS with each other. 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 MS and MG isolation and bacteria DNA were extracted by phenol/chloroform. The conserved region of 16S rRNA gene was applied for the detection of Mycoplasma genus in 163bp fragment and MG in 183 bp fragment and vlhA gene for detection of MS in 350 bp fragment. Then duplex PCR amplified the conserved region of 16S rRNA and vlhA genes was applied for detection of MG and MS. Twenty samples in Mycoplasma genus, 7 samples in MG and MS were positive in the single PCR whereas 3 samples simultaneously MG and MS were positive in the duplex PCR method. The results showed that duplex PCR was successful to simultaneous identification of MG and MS and suggested that duplex PCR is more rapid and inexpensive method than the single PCR for detection of MG and MS.