Protocol for high-throughput reservoir quantification across global HIV subtypes using a cross-subtype intact proviral DNA assay.

Autor:	Fish CS; Division of Human Biology, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA., Cassidy NAJ; Division of Human Biology, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA., Levy CN; Department of Obstetrics and Gynecology, University of Washington, Seattle, WA 98109, USA., Hughes SM; Department of Obstetrics and Gynecology, University of Washington, Seattle, WA 98109, USA., Jerome KR; Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA 8109, USA., Overbaugh J; Division of Human Biology, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA., Hladik F; Department of Obstetrics and Gynecology, University of Washington, Seattle, WA 98109, USA; Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Department of Medicine, University of Washington, Seattle, WA 98109, USA., Lehman DA; Division of Human Biology, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Department of Global Health, University of Washington, Seattle, WA 98109, USA. Electronic address: dlehman@fredhutch.org.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2022 Dec 16; Vol. 3 (4), pp. 101681. Date of Electronic Publication: 2022 Sep 28.
DOI:	10.1016/j.xpro.2022.101681
Abstrakt:	The cross-subtype intact proviral DNA assay (CS-IPDA) is a high-throughput method to quantify HIV reservoir size in populations infected with any of the dominant global HIV-1 subtypes. Our protocol includes genomic DNA isolation optimized to minimize DNA shearing, a reference droplet digital PCR (ddPCR) assay to quantify T cells and assess DNA shearing, and a multiplex ddPCR targeting three distinct regions across the HIV genome to quantify intact proviruses as an estimate of replication-competent proviruses in the reservoir. For complete details on the use and execution of this protocol, please refer to Cassidy et al. (2022). Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu