Molecular Identification of Mycoplasma agalactiae in Iran Based on P۳۰ Gene

Mycoplasma agalactiae (M. agalactiae) is known as the main etiological agent of contagious agalactia (CA). The CA is a disease affecting dairy sheep and goats, the main characteristics of which include keratoconjunctivitis, arthritis, and mastitis. This pathogen results in milk production reduction and suppression, thereby leading to serious economic loss. In the present study, ۱۲۵ sheep and goat samples were collected from ۱۵ provinces of Iran. Cultural and molecular methods were used for sample characterization. After extracting genomic DNAs using the phenol/chloroform method, the PCR technique was employed to detect Mycoplasma genus in ۱۶۳bp fragment of ۱۶S rRNA gene (M-PCR) and M. agalactiae in ۸۰۰bp fragment of conserve and specific P۳۰ lipoprotein gene (P۳۰-PCR) in cultural and clinical samples. Finally, to validate the experimental approach, a ۳۷۵ bp amplicon of P۸۰ lipoprotein was amplified using the MA-PCR. Out of ۱۲۵ samples under investigation, ۴۳ cases were positive, and Mycoplasma colonies were observed in the pleuropneumonia-like organisms agar culture. Based on the results of the M-PCR method, ۶۱ specimens (out of ۱۲۵ samples) were scored positive for Mycoplasma presence. Furthermore, ۲۰ samples were positive according to the P۳۰-PCR data. It should be mentioned that the MA-PCR was performed based on the P۸۰ gene on ۱۲۵ total samples to furtherverify the results for M.agalactiae detection. Based on the obtained data, P۳۰ and P۸۰ genes were presented and amplified in all Iranian M. agalactiae isolates (n=۲۰). Our results indicated that the P۳۰ gene was conserved and specific to all Iranian M. agalactiae isolates and this new P۳۰-PCR method (as an MA-PCR technique) might be useful in the detection of this pathogen.

کلیدواژه ها:

Contagious agalactia ، Culture ، Mycoplasma agalactiae ، PCR ، P۳۰ gene