Development Of Recombinant Antigens For Diagnosis Of Avian Mycoplasmosis

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dc.contributor.advisorRamadass, P
dc.contributor.advisorRaj, G. Dinakar
dc.contributor.advisorPrabhakar, T.G.
dc.contributor.advisorGanesan, P.I.
dc.contributor.authorVenkatesh, G
dc.contributor.authorTANUVAS
dc.date.accessioned2016-05-27T15:11:45Z
dc.date.available2016-05-27T15:11:45Z
dc.date.issued2006
dc.description.abstractAvian Mycoplasmosis is widespread and is one of the most common respiratory diseases of poultry causing significant economic losses in poultry industry. The disease occurs when birds infected with Mycoplasma gallisepticum (MG) or Mycoplasma synoviae (MS) are stressed. An attempt has been made in this study to express the antigenic proteins of MG and MS in E.coli and use them in diagnostic assay. Primers were designed with built in restriction enzyme sites for amplification of p75 and p29 (TM-1) genes of MG PG31, PCR amplification of the genes was done, cloned and sequenced. The genes were cloned into expression vector pPROEx HTb and expressed in E. coli DH5α. The full length vlhA gene of local field isolate MS427 was amplified using published primers and the 5’ end corresponding to MSPB protein sequences were sequenced. Based on the sequence, primers were synthesized, amplified , a smaller part of MSPB (MSPB-I) and a larger part of MSPB (MSPB-II) gene was cloned in the expression vector pPROEx HTb and expressed in E. coli DH5α.The recombinant proteins were purified using Ni-NTA affinity chromatography.The four recombinant proteins obtained in this study were characterized and evaluated as diagnostic antigen. The recombinant P29 of MG and recombinant MSPB-II protein of MS showed a potential to be used as diagnostic antigens. A highly sensitive indirect ELISA, a rapid simple easy to perform latex agglutination test and flow-through enzyme immunoassay were developed to diagnose MG and MS separately using their respective recombinant proteins. The MG and MS indirect ELISA had perfect agreement with commercial ELISA kit used in this study. The latex agglutination tests and flow-through enzyme immunoassays had substantial agreement with commercial kit.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/66397
dc.language.isoenen_US
dc.publisherTamil Nadu Veterinary and Animal Sciences Universityen_US
dc.subAnimal Biotechnology
dc.subjectAvian Mycoplasmosisen_US
dc.subjectMycoplasma gallisepticumen_US
dc.subjectMycoplasma synoviaeen_US
dc.subjectCloningen_US
dc.subjectRecombinant expressionen_US
dc.subjectELISAen_US
dc.subjectLatex agglutination testen_US
dc.subjectFlow-through enzyme immunoassayen_US
dc.these.typePh.D
dc.titleDevelopment Of Recombinant Antigens For Diagnosis Of Avian Mycoplasmosisen_US
dc.typeThesisen_US
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