Impact of naturally occurring amino acid variations on the detection of HIV-1 p24 in diagnostic antigen tests.

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_53B70E195427
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Impact of naturally occurring amino acid variations on the detection of HIV-1 p24 in diagnostic antigen tests.
Journal
BMC infectious diseases
Author(s)
Vetter B.N., Orlowski V., Niederhauser C., Walter L., Schüpbach J.
ISSN
1471-2334 (Electronic)
ISSN-L
1471-2334
Publication state
Published
Issued date
29/10/2015
Peer-reviewed
Oui
Volume
15
Pages
468
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
The detection of HIV-1 p24 antigen in diagnostic tests relies on antibodies binding to conserved areas of the protein to cover the full range of HIV-1 subtypes. Using a panel of 43 different virus-like particles (VLPs) expressing Gag from clinical HIV-1 isolates, we previously found that some highly sensitive tests completely failed to detect p24 of certain VLPs, seemingly unrelated to their subtype. Here we aimed to investigate the reason for this failure, hypothesising that it might be due to single amino acid variations in conserved epitopes.
Using amino acid alignment, we identified single amino acid variations at position 16 or 170 of p24, unique to those VLPs that failed to be detected in certain diagnostic tests. Through DNA-mutagenesis, these amino acids were changed to ones more commonly found at these positions. The impact of these changes on p24 detection was tested in commercial diagnostic tests as well as by Western Blot and ELISA, using epitope-specific antibodies.
Changing positions 16 or 170 to consensus amino acids restored the detection of p24 by the investigated diagnostic tests as well as by epitope-specific antibodies in Western Blot and ELISA. Hence, single amino acid changes in conserved epitopes can lead to the failure of p24 detection and thus to false-negative results. To optimise HIV diagnostic tests, they should also be evaluated using isolates which harbour less-frequent epitope variants.

Keywords
AIDS Serodiagnosis/methods, Amino Acids/genetics, Antigens, Viral/immunology, Blotting, Western, Enzyme-Linked Immunosorbent Assay/methods, Epitopes/immunology, HIV Core Protein p24/analysis, HIV Core Protein p24/genetics, HIV Core Protein p24/immunology, HIV-1/immunology, HIV-1/isolation & purification, Humans, Mutagenesis, Site-Directed, Reagent Kits, Diagnostic
Pubmed
Web of science
Open Access
Yes
Create date
31/01/2018 12:30
Last modification date
21/08/2019 7:09
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