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. 2021 Oct 11;2(4):100885.
doi: 10.1016/j.xpro.2021.100885. eCollection 2021 Dec 17.

HIV reservoir quantification by five-target multiplex droplet digital PCR

Claire N Levy  1 , Sean M Hughes  1 , Pavitra Roychoudhury  2 , Chelsea Amstuz  2 , Haiying Zhu  3 , Meei-Li Huang  3 , Dara A Lehman  4   5 , Keith R Jerome  2   3   5 , Florian Hladik  1   2   6
Affiliations

HIV reservoir quantification by five-target multiplex droplet digital PCR

Claire N Levy et al. STAR Protoc. .

Abstract

Most latent human immunodeficiency virus (HIV) proviruses are defective and cannot produce infectious virions. Thus, the number of HIV proviruses with intact genomes is a relevant clinical parameter to assess therapies for HIV cure. We describe high-molecular-weight DNA isolation, followed by restriction enzyme fragmentation that limits cutting within the HIV genome. Multiplexed droplet digital PCR quantifies five targets spanning the HIV genome to estimate potentially intact proviral copies. A reference assay counts the number of T lymphocytes and assesses the level of DNA shearing. For complete details on the use and execution of this protocol, please refer to Levy et al. (2021).

Keywords: Bioinformatics; Immunology; Microbiology; Molecular Biology.

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Conflict of interest statement

The authors declare no competing interests.

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