Detection of Mycoplasma gallisepticum by 16S rRNA PCR with specific primers in clinical samples

Subject Areas : Veterinary Clinical Pathology

S.A Pourbakhsh ¹ (Department of Microbiology, Faculty of Specialized Veterinary Sciences, Islamic Azad University, Science & Research Branch, Tehran, Iran)
افشین Zakeri ² (Ph.D Student of Poultry Diseases, Faculty of Specialized Veterinary Sciences, Islamic Azad University, Science & Research Branch, Tehran,)
نریمان Sheikhi ³ (Department of Clinical Science, Faculty of Specialized Veterinary Sciences, Islamic Azad University, Science & Research Branch, Tehran, Iran)
سعید Charkhkar ⁴ (Department of Clinical Science, Faculty of Specialized Veterinary Sciences, Islamic Azad University, Science & Research Branch, Tehran, Iran)
عباس Ashtari ⁵ (Mycoplasma Reference Lab. of Razi Vaccine and Serum Research Institute, Karadj, Iran)

Received: 2009-06-06 Accepted : 2010-01-04 Published : 2009-08-23

Keywords: PCR, Clinical samples, Mycoplasma Gallisepticum,

Abstract :

Mycoplasma gallisepticum (MG) as one the major pathogens of birds, causes significant economic losses in poultry industry. The main purpose of the present study was to detect Mycoplasma gallisepticum in clinical samples using the 16S rRNA PCR method. For serological screaming test, 18 commercial laying farms and 8 broiler breeder farms were selected and rapid serum agglutination test (RSAT) was performed. For polymerase chain reaction sampling, 10 of the 17 farms that were positive in RSAT were selected and 109 sterile swab samples were collected from the palatine cleft, trachea, air sacs and lungs in each farm. Three swabs from three birds were placed in test tube containing 1 ml of phosphate buffered saline and transferred to laboratory form PCR testing. In this study, specific primers for 16S rRNA gene were used. The aforementioned primers are totally specific for MG and can be differentiated from other Mycoplasmaand bacteria present in the trachea of poultry of the 26 farms examined, 17 farms were positive in RSAT serologic test. The 530 bp PCR product produced by specific primers of all field strains appeared on electrophoresis gel in 46 samples from 10 farms accounting to 42.2%.The 16S rRNA PCR with very high sensitivity can be employed in definitive diagnosis of Mycoplasma gallisepticum infection in vitro.  

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