ISSN:
1432-1440
Keywords:
AIDS (acquired immunodeficiency syndrome)
;
Expression cloning
;
HIV-1 (human immunodeficiency virus type 1)
;
Immunofluorescence
;
Recombinant protein
;
Westernblot
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
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 β-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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01726323
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