P. N. Dwivedi

Multiplex real-time PCR for identification of canine parvovirus antigenic types.

Canine parvovirus (CPV) is an important disease causing gastroenteritis and/or haemorrhagic gastroenteritis in dogs. There are four antigenic types of CPV reported worldwide viz. CPV 2, CPV 2a, CPV 2b and CPV 2c. The diagnosis of CPV with the identification of the antigen type responsible remains problematic. In the present study, identification as well as antigenic typing of CPV was done using a de novo multiplex real time PCR to combat the problem of antigenic type identification. From the study it could be concluded that the here developed multiplex real time PCR assay could be used for rapid detection of CPV as well as typing of its three antigenic types.

Isolation of Canine parvovirus with a view to identify the prevalent serotype on the basis of partial sequence analysis

Aim: The aim of this study was to isolate Canine parvovirus (CPV) from suspected dogs on madin darby canine kidney (MDCK) cell line and its confirmation by polymerase chain reaction (PCR) and nested PCR (NPCR). Further, VP2 gene of the CPV isolates was amplified and sequenced to determine prevailing antigenic type. Materials and Methods: A total of 60 rectal swabs were collected from dogs showing signs of gastroenteritis, processed and subjected to isolation in MDCK cell line. The samples showing cytopathic effects (CPE) were confirmed by PCR and NPCR. These samples were subjected to PCR for amplification of VP2 gene of CPV, sequenced and analyzed to study the prevailing antigenic types of CPV. Results: Out of the 60 samples subjected to isolation in MDCK cell line five samples showed CPE in the form of rounding of cells, clumping of cells and finally detachment of the cells. When these samples and the two commercially available vaccines were subjected to PCR for amplification of VP2 gene, a 1710 bp product was amplified. The sequence analysis revealed that the vaccines belonged to the CPV-2 type and the samples were of CPV-2b type. Conclusion: It can be concluded from the present study that out of a total of 60 samples 5 samples exhibited CPE as observed in MDCK cell line. Sequence analysis of the VP2 gene among the samples and vaccine strains revealed that samples belonged to CPV-2b type and vaccines belonging to CPV-2.

Detection and Isolation of Peste-des-petits-ruminants (PPR) Virus in Goats of Punjab Region (India)

Peste-des-petits-ruminants (PPR) is an acute highly contagious disease of sheep and goats causing high morbidity and mortality. PPR is endemic in India and outbreaks occur in sheep and goats regularly throughout the country leading to huge economic losses. The present study aimed at the detection of PPR virus in nasal and ocular swabs of goats from Punjab region and also to isolate the virus. For this purpose, 100 samples (50 ocular and 50 nasal swabs) were collected from 50 goats. The samples were subjected to sandwich Enzyme Linked Immunosorbent Assay and Reverse Transcriptase-Polymerase Chain Reaction for detection of PPR virus and also subjected to virus isolation in B95a cell line. The study revealed that PPR virus is present in the goat population in the region under study. Further, PPR virus could be successfully isolated in B95a cell line.