Optimization of variable number tandem repeat (VNTR) analysis in the classical PCR machines for typing of Burkholderia mallei
Subject Areas : Molecular Microbiology
Reza Najafpour
1
(M.Sc., Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord branch, Shahrekord, Iran.)
Nader Mosavari
2
(Assistance Professor, Razi Vaccine and Serum Research Institute, Karaj, Iran.)
Keyvan Tadayon
3
(Assistance Professor, Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord branch, Shahrekord, Iran.)
Elaheh Tajbakhsh
4
(Assistance Professor, Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord branch, Shahrekord, Iran.)
Keywords: Burkholderia mallei, BM140, BM1367, BM2065, BM2971,
Abstract :
Background & Objectives: 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 for 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 for the first time. Materials & Methods: In this study, B. mallei Razi 325, as a standard strain, was obtained from the microbial archive of the Razi Vaccine and Serum Research Institute. Following DNA extraction and PCR amplification of 4 loci, results of each locus was analysed independently and in different combinations. A special protocol was set in terms of temperature and concentrations of the cotenants (MgCl2 and primers) for each locus. Results: A comparison of the sequences of our isolate with the standard strains indicated the presence of polymorphism in these loci. Conclusion: Achievement of a common PCR protocol, which is able to proceed simultaneously four different reactions in one PCR run, was the primary outcome of this research.