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Szojka Z, Karlson S, Jansson M, et al. Quantification of HIV-2 DNA in Whole Blood. Bio Protoc. 2019;9(20):e3404doi: 10.21769/BioProtoc.3404.
Szojka, Z., Karlson, S., Jansson, M., & Medstrand, P. (2019). Quantification of HIV-2 DNA in Whole Blood. Bio-protocol, 9(20), e3404. https://doi.org/10.21769/BioProtoc.3404
Szojka, Zsófia, et al. "Quantification of HIV-2 DNA in Whole Blood." Bio-protocol vol. 9,20 (2019): e3404. doi: https://doi.org/10.21769/BioProtoc.3404
Szojka Z, Karlson S, Jansson M, Medstrand P. Quantification of HIV-2 DNA in Whole Blood. Bio Protoc. 2019 Oct 20;9(20):e3404. doi: 10.21769/BioProtoc.3404. eCollection 2019 Oct 20. PMID: 33654905; PMCID: PMC7853964.
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Bio Protoc. 2019 Oct 20;9(20):e3404. doi: 10.21769/BioProtoc.3404. eCollection 2019 Oct 20.

Quantification of HIV-2 DNA in Whole Blood.

Bio-protocol

Zsófia Szojka, Sara Karlson, Marianne Jansson, Patrik Medstrand

Affiliations

  1. Department of Translational Medicine, Clinical Virology, Lund University, Malmö, Sweden.
  2. Laboratory of Retroviral Biochemistry, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
  3. Department of Laboratory Medicine, Lund University, Lund, Sweden.

PMID: 33654905 PMCID: PMC7853964 DOI: 10.21769/BioProtoc.3404

Abstract

Time to AIDS infection is longer with HIV-2, compared to HIV-1, but without antiretroviral therapy both infections will cause AIDS-related mortality. In HIV-2 infection, monitoring of antiretroviral treatment (ART) efficacy is challenging since a large proportion of HIV-2-infected individuals displays low or undetectable plasma RNA levels. Hence, quantification of cellular DNA load may constitute an alternative method for monitoring ART efficacy. Moreover, sensitive HIV-2 DNA quantification protocols are also important for the characterization of the HIV-2 reservoirs, and ultimately for the development of HIV-2 cure strategies. We have developed a sensitive and robust HIV-2 DNA quantification protocol based on whole blood as DNA source, including normalization of leukocyte cell numbers using parallel quantification of the single copy porphobilinogen deaminase gene. The specificity and sensitivity of the assay was 100%. The limit of detection was 1 copy and limit of quantification was 5 copies. When applying this protocol to HIV-2 infected, it was found that HIV-2 viral DNA was detectable in individuals in whom viral RNA was undetectable or under quantification level. Thus, this method provides a sensitive approach to HIV-2 DNA viral quantification from whole blood of HIV-2 infected patients.

Copyright © 2019 The Authors; exclusive licensee Bio-protocol LLC.

Keywords: DNA load; HIV-2; Quantification; Whole Blood; qPCR

Conflict of interest statement

Competing interestsThe funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Authors have no conflict of interest.

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