Journal article
Adaptation of the intact proviral DNA assay to a nanowell-based digital PCR platform
C Tumpach, CR Cochrane, Y Kim, J Ong, A Rhodes, TA Angelovich, MJ Churchill, SR Lewin, S Telwatte, M Roche
Journal of Virus Eradication | Published : 2023
Abstract
Quantification of intact proviruses is a critical measurement in HIV cure studies both in vitro and in vivo. The widely adopted ‘intact proviral DNA assay’ (IPDA), designed to discriminate and quantify genetically intact HIV proviruses based on detection of two HIV sequence-specific targets, was originally validated using Bio-Rad's droplet digital PCR technology (ddPCR). Despite its advantages, ddPCR is limited in multiplexing capability (two-channel) and is both labor- and time intensive. To overcome some of these limitations, we utilized a nanowell-based digital PCR platform (dPCR, QIAcuity from Qiagen) which is a fully automated system that partitions samples into nanowells rather than dr..
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Grants
Awarded by National Institutes of Health
Funding Acknowledgements
& nbsp;J-Lat 10.6 (ARP-9849) and ACH2 (ARP-349) cells were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH. The authors acknowledge the award of a University of Melbourne Faculty of Medicine, Dentistry and Health Sciences Large Equipment Grant to SRL for purchase of the QIAcuity platform. The work was supported by the National Health and Medical Research Council (NHMRC) of Australia Ideas Grant (MJC and MR, #1183032) ;Practitioner Fellowship (SRL, #1135851) and Program Grant (SRL, #1149990) and from the National Institutes Allergy and Infectious Diseases Delaney AIDS Research Enterprise to find a cure (DARE) (SRL, 1UM1AI164560-01) . ST is supported by the Locarnini Fellowship in Virology at the Doherty Institute and The University of Melbourne Department of Infectious Diseases Research Support Package.