Impact of naturally occurring amino acid variations on the detection of HIV-1 p24 in diagnostic antigen tests.
Détails
Télécharger: 26511217_BIB_53B70E195427.pdf (660.07 [Ko])
Etat: Public
Version: Final published version
Licence: CC BY 4.0
Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_53B70E195427
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Impact of naturally occurring amino acid variations on the detection of HIV-1 p24 in diagnostic antigen tests.
Périodique
BMC infectious diseases
ISSN
1471-2334 (Electronic)
ISSN-L
1471-2334
Statut éditorial
Publié
Date de publication
29/10/2015
Peer-reviewed
Oui
Volume
15
Pages
468
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Publication Status: epublish
Résumé
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.
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.
Mots-clé
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
Oui
Création de la notice
31/01/2018 11:30
Dernière modification de la notice
21/08/2019 6:09