Molecular characterization of wild life and laboratory Burkholderia mallei isolates from Iran

Glanders caused by Burkholderia mallei, is a zoonotic disease. mallein test is the most used method in diagnosis of glanders. mallein is still prepared of standard B. mallei strain (s). In order to characterize the genomic properties, we have studied nucleotide structure of 23SrDNA, flip and BimA genes of a B.mallei strain that is used for preparation of mallein at Razi Institute and also a B.mallei isolate collected from a diseased tiger linked to the latest glanders outbreak at Tehran zoo. The isolated bacteria were cultivated on specific media. Bacterial suspensions were used to inoculate two guinea pigs in order to develop Strauss phenomenon. Genomic DNA was extracted by Isoamyl alcohol-Chloroform method. The PCR developed by using specific primers of B.mallei (three pairs) and common between B.mallei and B.pseoudomallei (one pair). The nucleotide sequence of the amplified genes was compared with standard fully sequenced genome available in gene bank. Both inoculated guinea pigs developed the characteristic Strauss phenomenon. Bacterial growth was the quantitative and qualitative different in the media and animal boudy. The amplificating four loci were successful. The nucleotide sequence analysis indicated these four loci were identical in B.mallei sterain/isolate invegtigented. This strategy represented a diagnostic algorithm suitable for differentiation between B.mallei and B.pseudomallei. Because importance of disease and limitation of mullein test in clinical and advanced cases of glanders using of sensitive and rapid diagnostic methods dependent on PCR and aknowledge of epidemiology molecular of glanders, is useful.