Development of Multiplex Polymerase Chain Reaction (mPCR): an emerging technique for the diagnosis of infectious organisms
Author(s) : Zaryyab Nayab, Aamir Ghafoor, Shahzada Khurram Syed, Muhammad Naeem Aamir & Naveed Munir
Abstract:
DOI: https://doi.org/10.52587/njhms.v5i2.80Objective: The health status of the community markedly depends upon the healthy livestock industry. This does not only contribute major role in the economic but also in health of human being worldwide. In the One Health perspective, the current research work was designed to develop four (04) sets of multiplex polymerase chain reactions (mPCR) for the concomitant recognition of bovine infections.
Material and Methods: The current research work was designed to develop four (04) sets of mPCR for the concomitant recognition of bovine abortive infections, including Salmonella spp. (mPCR1); Campylobacter fetus, Aspergillus spp. and Salmonella spp. (mPCR2); Salmonella spp., Campylobacter fetus, Brucella spp., Leptospira spp. and Listeria monocytogenes (mPCR3); Bovine viral diarrhoea virus (BVDV), Bluetongue virus and Bovine herpesvirus-1 (BoHV-1) (mPCR4). The procedure was evaluated on positive controls obtained commercially as well as from Microbiology Department of University of Veterinary and Animal Sciences, Lahore-Pakistan. These positive controls were optimized first by singlet PCR followed by multiplex PCR using specific forward and reverse primers.
Results: Results obtained from each multiplex PCR reaction were 100% compatible and in agreement with the respective PCR assays in which single genes were targeted.
Conclusion: It could be concluded that for the concomitant recognition and identification of the major bovine infectious abortive agents, these proposed sets of multiplex PCR assays will be more suitable which might help to proper timely management of such infectious diseases in the livestock an healthcare industry.
Keywords: Infectious diseases, Diagnostic techniques, multiplex PCR, Zoonosis