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Development and validation of HIV-1 Multiplex Serology.

L M Kranz1, B Gärtner2, A Michel3

  • 1Infections and Cancer Epidemiology, Infection, Inflammation and Cancer Research Program, German Cancer Research Center (DKFZ), Heidelberg, Germany; BioNTech Corporation, Mainz, Germany.

Journal of Immunological Methods
|January 21, 2019
PubMed
Summary

A new high-throughput HIV-1 Multiplex Serology test simultaneously detects antibodies to multiple human immunodeficiency virus-1 (HIV-1) antigens. This method accurately determines HIV-1 antibody status, aiding large-scale cancer development studies.

Keywords:
HIVHigh-throughputImmunoassayMultiplexSeroepidemiologySerology

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Area of Science:

  • Immunology
  • Virology
  • Epidemiology

Background:

  • Human immunodeficiency virus-1 (HIV-1) infection causes immunosuppression, increasing risks for opportunistic infections and cancers.
  • Accurate detection of HIV-1 antibody status is crucial for epidemiological studies, particularly concerning cancer development.
  • Conventional serology requires a two-step screening and confirmation process, limiting high-throughput applications.

Purpose of the Study:

  • To develop and validate a high-throughput multiplex serology assay for detecting HIV-1 antibody status.
  • To assess the utility of simultaneous detection of antibodies against multiple HIV-1 antigens for epidemiological research.
  • To evaluate the potential of identifying infection-associated antibody patterns for disease staging.

Main Methods:

  • Recombinant expression of seven HIV-1 antigens as glutathione-S-transferase (GST) fusion proteins in E. coli.
  • Immobilization of GST-fusion proteins onto distinct fluorescently coded bead sets for multiplexing.
  • Simultaneous detection of serum or plasma antibodies targeting multiple HIV-1 antigens in a suspension array.
  • Validation using 244 pre-determined reference sera, comparing results with conventional immunoassay methods.

Main Results:

  • The HIV-1 Multiplex Serology assay demonstrated high performance, comparable to conventional methods.
  • The multifunctional protein GAG was identified as a highly specific (99%) and sensitive (100%) marker for HIV-1 serostatus.
  • HIV-1 infected individuals recognized a median of 4 antigens, indicating a broad immune response, while negative sera recognized 0.
  • The assay eliminates the need for a two-step screening and confirmation approach.

Conclusions:

  • HIV-1 Multiplex Serology offers a validated, high-throughput alternative for determining HIV-1 antibody status.
  • This method facilitates large-scale epidemiological studies on HIV-1 and its association with cancer development.
  • The ability to detect antibodies against multiple antigens may aid in identifying patterns related to disease progression.