Detection of Nontuberculous Mycobacterial Species from Tissue Samples of Cattle and Buffaloes by PCR and PRA (PCR-RFLP)

Abstract

Background: Nontuberculous mycobacteria are opportunistic pathogens and some of them may cause disease in humans and animals causing pulmonary infections, mastitis, lesions in respiratory tract and lymph nodes of cattle, due to which they are being recognized worldwide and also interfere with the diagnosis of bovine tuberculosis. Methods: The present study was conducted for detection of nontuberculous mycobacterial species (NTM) in tissue samples (with and without tubercle lesions) in cattle and buffaloes from postmortem hall GADVASU, Ludhiana. Polymerase Chain Reaction and PCR-RFLP which involved hsp65 gene amplification (439 bp) and restriction analysis of amplified product was performed on 30 tissue samples for detection of nontuberculous mycobacterial species. Result: Three out of 30 samples showed hsp65 gene amplification and 2 were identified as M. kansasii using restriction analysis technique and one could not be identified as the RFLP patterns was different from other known PCR-RFLP profiles. NTM such as M. kansasi may cause infection in animals and PRA (PCR-Restriction Fragment Length Polymorphism Analysis) technique was found to be a rapid tool for identification and differentiation of NTM upto species level.