Standardization Of Indirect Enzyme Linked Immunosorbant Assay For Detection Of Antibodies Against Foot and Mouth Disease Virus Sdrotype "O"
By: Yasmeen Siddique
| Prof. Dr. Khushi Muhammad.
Contributor(s): Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: 
BookPublisher: 2005Subject(s): Department of MicrobiologyDDC classification: 0879,T Dissertation note: An indirect ELISA was standardized for titration of anti-FMD-serotype-specific antibodies. In this test polystyrene plates were coated with known FMD serotype "O" virus using carbonate/bicarbonate buffer. The blank spaces were blocked with horse serum. The immunoplate was coated with anti-FMD "O" virus specific serum from vaccinated calves. After washing, the plate was coated with rabbit anti-bovine-Ig-specific-antibodies-horse radish peroxidase conjugate. After washing, the plate was coated with HRP specific substrate. Development of color was recorded in form of OD value using ELISA reader. During the standardization of ELISA, flat bottom ELISA plates among all types of plates, 1:10 diluted virus among different dilutions of FMD "O" type virus, 1:10 diluted serum from buffalo calves vaccinated with FMD "O" type specific vaccine, 1:4000 dilution of conjugate and incubation of 4º C for coating the virus showed good results. In each experiment, plateau region, test back ground and plate back ground was recorded. Results of the study will help in establishment of an economical, sensitive, reliable indirect ELISA that subsequently be helpful to understand the percent prevalence of FMD serotypes in Pakistan and efficacy of FMD vaccines.
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UVAS Library Thesis Section | Veterinary Science | 0879,T (Browse shelf) | Available | 0879,T |
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An indirect ELISA was standardized for titration of anti-FMD-serotype-specific antibodies. In this test polystyrene plates were coated with known FMD serotype "O" virus using carbonate/bicarbonate buffer. The blank spaces were blocked with horse serum. The immunoplate was coated with anti-FMD "O" virus specific serum from vaccinated calves. After washing, the plate was coated with rabbit anti-bovine-Ig-specific-antibodies-horse radish peroxidase conjugate. After washing, the plate was coated with HRP specific substrate. Development of color was recorded in form of OD value using ELISA reader. During the standardization of ELISA, flat bottom ELISA plates among all types of plates, 1:10 diluted virus among different dilutions of FMD "O" type virus, 1:10 diluted serum from buffalo calves vaccinated with FMD "O" type specific vaccine, 1:4000 dilution of conjugate and incubation of 4º C for coating the virus showed good results. In each experiment, plateau region, test back ground and plate back ground was recorded. Results of the study will help in establishment of an economical, sensitive, reliable indirect ELISA that subsequently be helpful to understand the percent prevalence of FMD serotypes in Pakistan and efficacy of FMD vaccines.
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