Browsing by Author "Kapgate, Sunil"
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ThesisItem Restricted Development of non radioactive DNA probe for diagnosis of bovine herpes virus-1(LUVAS, 2006) Kapgate, Sunil; MinakshiBovine herpesvirus-l (BHV-1), a member of the family Herpesviridae and subfamily Alphaherpesvirinae is an important cause of respiratory and genital diseases in cattle. Infection with BHV-1 occurs worldwide and causes serious economic losses due to mortality, abortions, decreased milk production and loss of weight. The current techniques for detection of BHV- 1 in the diagnostic laboratories include virus isolation, and antigen detection by ELISA. These techniques lack sensitivity and are both time consuming and expensive. Keeping this point in view the present study was undertaken to develop and evaluate the non radioactive probe for detection of BHV-1. The non radioactive DNA probe was successfully evaluated for detection of BHV-1 in biological field samples. By using DNA probe total a 4 Nasal swabs, 8 vaginal swabs, lung, liver and cotyledons of aborted material were detected positive. Efficiency of two different primer pairs belonging to gI and gB glycoproteins, for detection of BHV-1 was compared. The gI specific PCR detected upto 1fg of viral DNA from purified BHV-1. Whereas the gB gene specific PCR detected upto 10fg of nucleic acid The gI gene specific PCR was used to confirm the results of probe hybridization assay. Probe positive all the samples also detected positive by PCR. None of the probe negative samples was found positive by PCR. The techniques could be useful for screening of large number of biological samples. The chelex method of DNA isolation from clinical samples included a fast, safe and convenient extraction procedure. The study clearly indicated that, the novel extraction method with non radioactive DNA probe hybridization assay provide simple, rapid and sensitive diagnostic tool for detection of BHV-1 infection in biological samples especially when large number have to be processed.ArticleItem Open Access Genotypic and Pathotypic Characterization of Newcastle Disease Viruses from India(PLOS ONE, 2011-12) Tirumurugaan, K.G.; Kapgate, Sunil; Vinupriya, Manavalan K.; Vijayarani, K; Kumanan, K; Elankumaran, Subbiah; TANUVASNewcastle disease virus (NDV) is an avian paramyxovirus that causes significant economic losses to the poultry industry in most parts of the world. The susceptibility of a wide variety of avian species coupled with synanthropic bird reservoirs has contributed to the vast genomic diversity of this virus as well as diagnostic failures. Since the first panzootic in 1926, Newcastle disease (ND) became enzootic in India with recurrent outbreaks in multiple avian species. The genetic characteristics of circulating strains in India, however, are largely unknown. To understand the nature of NDV genotypes in India, we characterized two representative strains isolated 13 years apart from a chicken and a pigeon by complete genome sequence analysis and pathotyping. The viruses were characterized as velogenic by pathogenicity indices devised to distinguish these strains. The genome length was 15,186 nucleotides (nt) and consisted of six non-overlapping genes, with conserved and complementary 39 leader and 59 trailer regions, conserved gene starts, gene stops, and intergenic sequences similar to those in avian paramyxovirus 1 (APMV-1) strains. Matrix gene sequence analysis grouped the pigeon isolate with APMV-1 strains. Phylogeny based on the fusion (F), and hemagglutinin (HN) genes and complete genome sequence grouped these viruses into genotype IV. Genotype IV strains are considered to have ‘‘died out’’ after the first panzootic (1926–1960) of ND. But, our results suggest that there is persistence of genotype IV strains in India.