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Publisher: PVNR TVU
Type: Thesis
Series/Report no.: D;415
Agrotags: genotypes, planting, grain, developmental stages, yields, rice, genetics, productivity, seasons, diseases
Abstract: Goat pox is a highly contagious disease caused by goatpox virus which belongs to the genus Capripoxvirus of the family Poxviridae. The prevalence of this disease among small ruminants is rampant and the economic losses from the disease are considerably high posing serious threat to global trade of the animals and their products. Though various tests have been demonstrated for the serological diagnosis of the disease they are time consuming and not readily available. Towards achieving this goal,a rapid and convenient sero-diagnostic technique like Indirect Enzyme Linked immunosorbent assay was attempted and developed to detect goat pox virus antibodies more effectively.An attempt was also made to differentiate sheep pox and goat pox viruses using polymerase chain reaction. Name of the Author : LAXMI ROHINI MADUPU Title of the thesis : CHARACTERISATION OF GOAT POX VIRUS Degree to which it is submitted : MASTEROF VETERINARY SCIENCE Faculty : VETERINARY SCIENCE Discipline : MICROBIOLOGY Major Advisor : Dr. K. DHANA LAKSHMI University : SRI VENKATESWARA VETERINARY UNIVERSITY Year of Submission : 2015 Goat pox virus successfully propagated in vero cells was used for the purification of virus by sucrose density gradient centrifugation. The virus was concentrated as aclear opalescent band at interface of 50% and 60% sucrose density gradients. Purified protein analysis was done by SDS-PAGE for the visualization of viral polypeptides. Five major polypeptides specific for Capripoxviruses with molecular weights ranging from 20k Da to 67k Da along with some comigrating bands were observed. The purified virus was used as an antigen for the development and standardization of indirect ELISA.Optimization of anti-goat HRPO conjugate, concentration of antigen and serum dilutions was done by checker board titration. It was found that 1:10,000 dilution of conjugate, 1:100 dilution of antigen (549.20μg/ml) per well and 1:100 of dilution of serum samples were optimum for the detection of antibodies to goat pox virus. Hyper immune serum was raised in goat and sheep against goat pox and sheep pox respectively to test for detection of antibodies using indirect ELISA and SNT. Indirect ELISA was compared with serum neutralization test (SNT) for the detection of antibodies using polyclonal sera. It was found that the relative sensitivity and specificity of ELISA and SNT was 85.5% and 39.6% respectively. Indirect ELISA was found to be more acceptable, rapid, inexpensive and sensitive than SNT to monitor the immune status of vaccinated goats.Cross reaction studies were conducted between sheep pox and goat pox viruses using indirect ELISA which proved that the two viruses are serologically indistinguishable. Polymerase chain reaction (PCR) was used for laboratory confirmation of the virus in field samples received from different outbreaks using capripox specific primers. RPO30 gene specific primers were used for the differentiation of sheep pox and goat pox viruses by PCR based on a 21- nucleotide deletion exclusively in SPV resulting in difference in size of the amplicons of SPV and GPV. This method of differentiation is simple and rapid for improved diagnosis of capripox infections in mixed populations of sheep and goat
Subject: Veterinary Microbiology
These Type: M.SC
Issue Date: 2015-01
Appears in Collections:Thesis

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