A crude extract of axenically grown amoebae was used as antigen in order to develop an AB microELISA for the detection and quantitation of E. histolytica-specific IgG antibodies. This ELISA was used to screen individual sera of patients suffering from invasive amoebiasis (n=47)and control individuals(n=33). Significant titers of E-histolytica- specific IgG antibodies were present only in sera of patients suffering from invasive amoebiasis. The AB-microELISA had a sensitivity of 98% and a specificity of 97%.
Immunoblot analysis of the crude E. histolytica extract indicated the presence of several antigenic proteins. One of the common antigenic proteins recognized by the individual patients' sera had a molecular weight of 160.170 kDa Sera of five patienta with high titers of E. histotytica-specific IgG antibodies were used to prepare a serum pool. This pooled serum was used for immunoscreening of an E. histolyitca cDNA expression library prepared in the phage vector l ZAP-II. A strongly immunoreactive phage done was identified, from which he recombinant phagemid was released by in vivo excision for characterization of the cDNA insert.
| Identifer | oai:union.ndltd.org:IISc/oai:etd.ncsi.iisc.ernet.in:2005/112 |
| Date | 08 1900 |
| Creators | Shenai, Bhaskar R |
| Contributors | Subba Rao, P V |
| Publisher | Indian Institute of Science |
| Source Sets | India Institute of Science |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis and Dissertation |
| Format | 4611050 bytes, application/pdf |
| Rights | I grant Indian Institute of Science the right to archive and to make available my thesis or dissertation in whole or in part in all forms of media, now hereafter known. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. |
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