Optimization of variable number tandem repeat (VNTR) analysis in the classical PCR machines for the typing of Burkholderia mallei

Glander is one of the important zoonotic diseases that is usually caused in solipeds including horse, donkey, mule, zebra and rarely in human. In the recent years, many molecular methods have been developed in the genome study of the Burkholderia mallei. The present study was aimed to optimize VNTR method on the basis of BM140, BM1367, BM2065 and BM2971 loci in order to molecular typing of B. mallei. In this study, B. mallei Razi325 (Razi Type Culture Collection RTCC: 2375) as the standard strain was obtained from the microorganisms archive of the Razi Vaccine and Serum Research Institute. The specific primers were designed for the amplification of fragments in the ranges of 450 to 750 base pairs. The comparision of the sequences between theour isolate and standard strains indicated polymorphism in the 4 loci. The major result of this research is developing of the unit PCR protocol for amplification of the fragments.