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

Autor: V. Orlowski, Louise Walter, Jörg Schüpbach, Christoph Niederhauser, Beatrice N. Vetter
Přispěvatelé: University of Zurich, Vetter, Beatrice N
Rok vydání: 2015
Předmět:
10028 Institute of Medical Virology
viruses
Blotting
Western
HIV Core Protein p24
Human immunodeficiency virus (HIV)
610 Medicine & health
Enzyme-Linked Immunosorbent Assay
medicine.disease_cause
Epitope
Epitopes
Western blot
Antigen
medicine
Humans
Amino Acids
Antigens
Viral
chemistry.chemical_classification
biology
medicine.diagnostic_test
HIV
AIDS Serodiagnosis
virus diseases
Diagnostic test
2725 Infectious Diseases
AIDS Serodiagnosis/methods
Amino Acids/genetics
Antigens
Viral/immunology
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
Mutagenesis
Site-Directed
Reagent Kits
Diagnostic
p24
Virology
Amino acid
Detection
Escape
Infectious Diseases
chemistry
Parasitology
Diagnostic tests
HIV-1
biology.protein
570 Life sciences
Antibody
Research Article
Zdroj: BMC Infectious Diseases
BMC infectious diseases, vol. 15, pp. 468
ISSN: 1471-2334
DOI: 10.1186/s12879-015-1174-7
Popis: Background 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. Methods 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. Results and Conclusions 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.
Databáze: OpenAIRE
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