• Login
    View Item 
    •   ResearchSpace Home
    • College of Agriculture, Engineering and Science
    • School of Life Sciences
    • Biotechnology
    • Microbiology
    • Masters Degrees (Microbiology)
    • View Item
    •   ResearchSpace Home
    • College of Agriculture, Engineering and Science
    • School of Life Sciences
    • Biotechnology
    • Microbiology
    • Masters Degrees (Microbiology)
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    The develolpment of a rapid diagnostic test for the detection of haemophilus ducrey.

    Thumbnail
    View/Open
    Thesis. (17.62Mb)
    Date
    2010
    Author
    Pillay, Mona.
    Metadata
    Show full item record
    Abstract
    Aim: To develop an antigen detection test that would quickly exclude H. ducreyi infection in individuals with genital ulcers. Materials and Methods: H. ducreyi strains A54 and A68 were grown on Modified Bieling (MB) agar plates and in MB broth under microaerophilic conditions. The 58.5 kDa GroEL Heat Shock Protein (HSP) was extracted from H. ducreyi strain A54 by means of sonication. The purified HSP was used to raise antibodies in rabbits. HSP determination and separation was done on SDS PAGE gels and protein was eluted by means of a passive elution process. Antibody was purified by affinity chromatography and a fraction of the antibody was conjugated to a chromogen to be used as a detection antibody. An ELISA was developed to evaluate the antibody response to the HSP. A second ELISA was developed to evaluate test parameters. Results: A good immune response was achieved with the crude serum of one of the three rabbits when tested against the antigen by means of ELISA. However, after purification of the IgG from the serum of the same rabbit no antigen-antibody binding was observed. Anti-rabbit IgG was able to recognise the antibodies. Discussion and Conclusion: While the Fc portion of the purified IgG remained active, the Fab portion of the antibody had lost biological activity. This loss of biological activity of antibody can be attributed to the low pH of the elution buffers used during the purification steps. Alternative antibody purification systems need to be explored. The use of monoclonal antibodies also needs to be considered.
    URI
    http://hdl.handle.net/10413/9565
    Collections
    • Masters Degrees (Microbiology) [91]

    DSpace software copyright © 2002-2013  Duraspace
    Contact Us | Send Feedback
    Theme by 
    @mire NV
     

     

    Browse

    All of ResearchSpaceCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsAdvisorsTypeThis CollectionBy Issue DateAuthorsTitlesSubjectsAdvisorsType

    My Account

    LoginRegister

    DSpace software copyright © 2002-2013  Duraspace
    Contact Us | Send Feedback
    Theme by 
    @mire NV