ASSESSMENT OF GENETIC PATTERN MYCOPLASMA AGALACTIAE ISOLATED FROM EAST AZARBAIJAN AND ELAM BY METHOD OF MLST TYPING

Background and Aim:Mycoplasma agalactiae is the main etiologic agent of contagious agalactia, a serious syndrome affecting sheep and goat associated with several clinical signs, including mastitis, arthritis, keratoconjunctivitis, pneumonia and septicaemia. The aim of this study was conducted to genotype M.agalactiae strains isolated from East Azarbaijan and Elam in milk samples using Multi Locus Sequence Typing (MLST) method.Methods:To perform MLST analysis bacterial DNA was extracted by simple boiling method, then primers were designed based on the sequence proposed by the MLST database (https://pubmlst.org/magalactiae/). The PCR products obtained from amplification of five housekeeping genes (dnaA, gltX , gyrB , metS, tufA) were sequenced. The nucleotide sequences of five genes in Target isolate were analyzed in the MLST database, then by using characterization of the alleles specific to each gene, the sequence types (ST) of all isolates were determined.Results:In consequence, at dnaA, gltX, gyrB, metS and tufA loci of all the target strains alleles 1, 21, 2, 2 and 1 were identified, respectively. A new sequence type 34 (ST34) was identified from archived target isolates at Razi institute for the first time in Iran.Conclusion:MLST method is portable, rapid, and very stable and is useful tool for epidemiological studies. We recommend using MLST for genotyping the isolates of other endemic areas of the country.

کلیدواژه ها:

Mycoplasma agalactiae ، MLST ، PCR ، strain