Serologische AIDS-Diagnostik mit gentechnisch gewonnenen Polypeptiden des menschlichen Immundefizienz-Virus (HIV-1) |
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Authors: | A Lenz J von Hintzenstern O Erlwein S Ellinger M Bröker B Fleckenstein G Jahn |
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Institution: | (1) Institut für Klinische und Molekulare Virologie der Universität Erlangen-Nürnberg, Germany;(2) Behringwerke AG, Marburg |
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Abstract: | Summary Serologic testing for human immunodeficiency virus type 1 (HIV-1) is currently based on enzyme linked immunosorbent assay (ELISA) as screening method. Positive ELISA-results have to be confirmed by at least one second procedure such as Western blotting or immunofluorescence. To obtain new diagnostic reagents for confirmatory testing, we expressed viral antigens in procaryotic systems. Peptides representing epitopes of structural core (gag)- and envelope (env)-proteins of HIV were produced inE. coli as stable immunogenic -galactosidase fusion proteins. Recombinant proteins were taken for immunoblot-assays. The results of Western blotting with those fusion proteins were in general comparable with conventional ELISA, immunofluorescence, immunoblot with cell-culture derived virus and commercially available ELISA tests based on recombinant proteins. Immunoblots using recombinant transmembrane protein (gp41) derived polypeptide were more sensitive than the conventional procedure with purified virion proteins. Western blotting with recombinant fusionproteins provide reliable and inexpensive serodiagnostics without handling of infectious cell cultures. Abkürzungsverzeichnis core- (gag-) Proteine
Innere Virus-Strukturproteine
- HIV-I
Humanes Immundefizienz-Virus Typ 1
- ELISA
Enzyme linked immunosorbent assay
- enve- lope- (env-) Proteine
Äußere Virus-Hüllproteine
- gp 41
Glykoprotein 41
- p 24
Protein 24 |
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Keywords: | AIDS (acquired immunodeficiency syndrome) Expression cloning HIV-1 (human immunodeficiency virus type 1) Immunofluorescence Recombinant protein Westernblot |
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