Please use this identifier to cite or link to this item: http://krishikosh.egranth.ac.in/handle/1/5810026416
Authors: Thomas, Jijo V.
Advisor: Minakshi
Title: Molecular characterization and complete genome sequencing of field isolates of canine parvovirus
Publisher: LUVAS
Language: en
Type: Thesis
Keywords: Canine parvovirus, Genotype, PCR-RFLP, Sequencing
Abstract: The present study was conducted to detect canine parvovirus in fecal samples of diarrheic dogs, to determine which genotype was present predominantly in the field and sequencing of the field isolate. VP2 gene based semi nested polymerase chain reaction was used to detect canine parvovirus in the field isolate and some virus samples were adapted in the CRFK cell line. Genotype specific primers are used to differentiate the field isolate into CPV2a and CPV2b.PCR RFLP was used to differentiate the field strain from the vaccine strain. The whole genome sequencing was performed using self designed primers which will give overlapping sequences and the sequences generated were analyzed to obtain the phylogenetic tree. Out of 102 fecal samples collected, 44 were found to positive for canine parvovirus on the basis of VP2 gene based semi nested polymerase chain reaction. Out of the 44 positive samples, 31 were un vaccinated and 2 were vaccinated and for the rest status were not known. The study shows that dog owners are not vaccinating their dogs properly. The genotypic differentiation study has shown that 36 samples belong to CPV2b and 8 belong to CPV2a out of the 44 positive fecal samples. Thus CPV2b is found to be the predominant genotype circulating in Hisar region of the Haryana followed by CPV2a.The PCR- RFLP study was carried out in the 747bp VP2 region by using the enzyme RsaI. The RE analysis revealed that there is a significant difference between field strain and vaccine strain. The vaccine strain which consists of CPV2 strain has provided a different RE pattern. Since the predominant genotype running in Hisar were CPV2a and CPV2b, that vaccine strain cannot provide complete immunity against CPV in domestic dog population so the vaccine should be provided with the predominant genotype circulating locally. The isolation of virus in CRFK cell line is found to be successful as positive results were obtained on PCR. The isolation of virus in cell line is one of the method of detection of CPV and is found to be true. The phylogenetic tree constructed based on both the NS gene showed that the Hisar isolate is evolutionarily more closer to USA isolate but the two isolates are grouped in separate clads. The phylogenetic tree constructed on the basis of VP gene showed that the Hisar isolate is more closer to Chinese isolate indicating that the two isolates are evolved from a common ancestor CPV. The present study showed that puppies within 6 weeks were affected more with CPV (70%) clearly substantiating that puppies are more prone to CPV infection.
Subject: Animal Biotechnology
Theme: Molecular characterization and complete genome sequencing of field isolates of canine parvovirus
These Type: M.V.Sc.
Issue Date: 2012
Appears in Collections:Thesis

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