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20101
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Subject: Veterinary Microbiology × Author: DEEPIKA KUMARI, GEDDADA × End date: 2018 × Start date: 2010 × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    CHARACTERIZATION OF CANINE PARVOVIRUS
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-08) DEEPIKA KUMARI, GEDDADA; DHANALAKSHMI, K(MAJOR); NARASIMHA REDDY, Y; MADHURI, D; REDDY, M.R
    ABSTRACT: The present study was taken up with a view to isolate canine parvovirus (CPV) from clinical cases and characterize them with regard to growth in cell culture, protein analysis and nucleic acid analysis. Further, haemagglutination of swine RBC, polymerase chain reaction and immunochromatography tests were compared for their efficacy in diagnosis of canine parvovirus infection. Ten faecal samples were collected from dogs suspected for canine parvovirus infections. The dogs exhibited symptoms like haemorrhagic enteritis, fever and vomition. The samples were tested for CPV antigen by haemagglutination of swine RBC as a preliminary test before attempts to isolate the virus. All the samples were positive with HA titres ranging from 32-1024. Attempts were made to isolate canine parvovirus from these faecal samples by passaging in CRFK and MDCK cells. Each of the sample was passaged ten times in both cell lines. There was no cytopathic effect in CRFK and MDCK cell lines at 10th passage. The presence of virus was demonstrated at different passage levels in CRFK but not in MDCK. However, the known isolate (IIL) caused focal rounding and aggregation of cells in CRFK but not in MDCK. With a view to characterize canine parvovirus, one isolate of canine parvovirus was purified employing sucrose density gradient centrifugation method. This method revealed purified virus as single light scattering band at 20% sucrose layer of the gradient. The purified virus gave one single precipitation line with hyperimmune serum in agar gel immunodiffusion test, confirming the isolate. The polypeptide analysis of the virus by SDS-PAGE revealed two polypeptide bands with molecular weights of 65kda and 62 kda. All the ten samples were positive for CPV by PCR employing the primer CPV-2ab which amplifies both 2a and 2b strains. However only one of these samples could be amplified by the primer CPV-2b specific for 2b strains of CPV. The known isolate (IIL) belonged to 2a strain. Three tests: Haemagglutination, PCR and rapid immunochromatographic tests (Rapigen kit test) were compared for their efficacy in the diagnosis of CPV. All ten samples were positive by haemagglutination test and PCR while only four samples were positive by rapigen kit indicating that rapigen kit can detect CPV only in faecal sample with HA titre of 512 and above. Further studies are required on more number of samples for studies on efficacy of diagnostic tests for canine parvovirus infection