Comparison of Detection Limits of Fourth- and Fifth-Generation Combination HIV Antigen-Antibody, p24 Antigen, and Viral Load Assays on Diverse HIV Isolates

J Clin Microbiol. 2018 Jul 26;56(8):e02045-17. doi: 10.1128/JCM.02045-17. Print 2018 Aug.

Abstract

Detection of acute HIV infection is critical for HIV public health and diagnostics. Clinical fourth-generation antigen (Ag)/antibody (Ab) combination (combo) and p24 Ag immunoassays have enhanced detection of acute infection compared to Ab-alone assays but require ongoing evaluation with currently circulating diverse subtypes. Genetically and geographically diverse HIV clinical isolates were used to assess clinical HIV diagnostic, blood screening, and next-generation assays. Three-hundred-member panels of 20 serially diluted well-characterized antibody-negative HIV isolates for which the researchers were blind to the results (blind panels) were distributed to manufacturers and end-user labs to assess the relative analytic sensitivity of currently approved and preapproved clinical HIV fourth-generation Ag/Ab combo or p24 Ag-alone immunoassays for the detection of diverse subtypes. The limits of detection (LODs) of virus were estimated for different subtypes relative to confirmed viral loads. Analysis of immunoassay sensitivity was benchmarked against confirmed viral load measurements on the blind panel. On the basis of the proportion of positive results on 300 observations, all Ag/Ab combo and standard sensitivity p24 Ag assays performed similarly and within half-log LODs, illustrating the similar breadth of reactivity and diagnostic utility. Ultrasensitive p24 Ag assays achieved dramatically increased sensitivities, while the rapid combo assays performed poorly. The similar performance of the different commercially available fourth-generation assays on diverse subtypes supports their use in broad geographic settings with locally circulating HIV clades and recombinant strains. Next-generation preclinical ultrasensitive p24 Ag assays achieved dramatically improved sensitivity, while rapid fourth-generation assays performed poorly for p24 Ag detection.

Keywords: Ag detection; Ag/Ab combo; HIV; analytic sensitivity; diagnostics; ultrasensitive Ag; viral load.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Serodiagnosis / methods*
  • AIDS Serodiagnosis / standards*
  • Benchmarking
  • HIV / immunology
  • HIV / isolation & purification*
  • HIV Antibodies / blood
  • HIV Antigens / blood
  • HIV Antigens / immunology
  • HIV Core Protein p24 / blood*
  • HIV Core Protein p24 / immunology*
  • HIV Infections / blood
  • HIV Infections / diagnosis*
  • Humans
  • Immunoassay / standards*
  • Limit of Detection
  • Sensitivity and Specificity
  • Viral Load / standards*

Substances

  • HIV Antibodies
  • HIV Antigens
  • HIV Core Protein p24