Vol. 7, Issue 11 (2018)

Author(s):
Athira S, Priya PM, Sheikh Moin Ahmad, Rinsha Balan and Mini M

Abstract:
Riemerella anatipestifer, E. coli and Salmonella are the three bacterial agents which produce concurrent infection in ducks. These agents produce more or less similar lesions, and hence differential diagnosis makes it difficult. Conventional cultural methods are laborious and time consuming, and not recommended at the time of outbreak. Molecular techniques like polymerase chain reaction (PCR) are widely employed for rapid identification of the etiological agents. Hence, a study was framed with an objective to develop multiplex Polymerase Chain Reaction (m-PCR) for the rapid detection and differential diagnosis of these bacterial agents. The positive isolates of R. anatipestifer, E. coli and Salmonella were revived and their DNA was extracted. After checking the purity and concentration, they were subjected to individual PCR for optimization of annealing temperature and primer concentration. Primer based on DnaB gene, PhoA gene and InvA gene were chosen to amplify the R. anatipestifer, E. coli and Salmonella genomic DNAs. Multiplex PCR was standardized in which the DNA of R. anatipestifer yielded an amplicon of 610 bp, whereas E. coli and Salmonella genomic DNA gave 720 bp and 256 bp product size. All the three primers were highly specific and no cross reactions were detected. Also they yield the products which are distinguishable. The developed m-PCR could be employed for the routine detection and differentiation R. anatipestifer, E. coli and Salmonella infection in ducks.