Molecular based diagnosis of rinderpest and peste des petits ruminants virus in Pakistan

Differential diagnosis of Rinderpest (RP) and Peste des petits ruminants (PR) by host species and clinical signs is not always exact; hence, laboratory confirmation is necessary to establish the cause of disease and to distinguish these two viruses, where small animals are source of infection for large ruminants. In this study, reverse transcription polymerase chain reaction (RT-PCR) was standardized against RP and PR. RNA of these viruses was isolated using denaturing solution (Solution D) containing guanidium thiocynate, sarcosyl, sodium citrate and beta-2 mercapto-ethanol. Regular PCR using morbilli specific oligonuclotides based on the sequences of conserved region of F and P-gene. These F and P gene primers amplified 429bp and 372bp, respectively for both RP and PPR. Nested PCR was performed to differentiate RP and PPR using F-gene amplicons by F3A/F4A and FlA/F2A primers, respectively. These sets of primers amplified a product of 235bp and 309bp for RP and PPR, respectively. The results of the study indicated that RT-PCR can be successfully used for detection and differential diagnosis of RP and PPR.