跳转至内容
Merck
CN
  • Impact of naturally occurring amino acid variations on the detection of HIV-1 p24 in diagnostic antigen tests.

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

BMC infectious diseases (2015-10-30)
Beatrice N Vetter, Vanessa Orlowski, Christoph Niederhauser, Louise Walter, Jörg Schüpbach
摘要

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.

材料
货号
品牌
产品描述

Sigma-Aldrich
硫酸, ACS reagent, 95.0-98.0%
Sigma-Aldrich
水, Nuclease-Free Water, for Molecular Biology
Sigma-Aldrich
水, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
水, Deionized
Sigma-Aldrich
硫酸, 99.999%
Sigma-Aldrich
水, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
硫酸, puriss. p.a., for determination of Hg, ACS reagent, reag. ISO, reag. Ph. Eur., 95.0-97.0%
Sigma-Aldrich
水, for molecular biology, sterile filtered
Sigma-Aldrich
3,3′,5,5′-四甲基联苯胺, ≥99%
Sigma-Aldrich
水, BioPerformance Certified
Sigma-Aldrich
3,3′,5,5′-四甲基联苯胺, ≥98% (TLC)
Sigma-Aldrich
硫酸, puriss., meets analytical specification of Ph. Eur., BP, 95-97%
Sigma-Aldrich
水, ACS reagent
Sigma-Aldrich
水, for cell biology, sterile ultrafiltered
Roche
逆转录酶检测,比色法, suitable for enzyme immunoassay, kit of 1 (14 components), sufficient for ≤200 tests
Sigma-Aldrich
3,3′,5,5′-四甲基联苯胺, ≥98.0% (NT)
Sigma-Aldrich
水, PCR Reagent
Sigma-Aldrich
3,3′,5,5′-四甲基联苯胺, tablet, 1 mg substrate per tablet
Sigma-Aldrich
E-Toxate 水, endotoxin, free