| Title: | An assessment of the reproducibility of reverse transcription digital PCR quantification of HIV-1 |
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| Authors: | ID Falak, Samreen (Author)
ID Macdonald, Rainer (Author)
ID Busby, Eloise J. (Author)
ID O'Sullivan, Denise M. (Author)
ID Milavec, Mojca (Author)
ID Plauth, Annabell (Author)
ID Kammel, Martin (Author)
ID Zeichhardt, Heinz (Author)
ID Grunert, Hans-Peter (Author)
ID Kummrow, Andreas (Author)
ID Huggett, Jim F. (Author) |
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| Files: |  URL - Source URL, visit https://doi.org/10.1016/j.ymeth.2021.03.006
 PDF - Presentation file, download (869,23 KB)
MD5: 59CF6A5B70A39431A8C01A99430F6BA8
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| Language: | English |
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| Typology: | 1.01 - Original Scientific Article |
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| Organization: |  NIB - National Institute of Biology
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| Abstract: | Viral load monitoring in human immunodeficiency virus type 1 (HIV-1) infection is often performed using reverse transcription quantitative PCR (RT-qPCR) to observe response to treatment and identify the development of resistance. Traceability is achieved using a calibration hierarchy traceable to the International Unit (IU). IU values are determined using consensus agreement derived from estimations by different laboratories. Such a consensus approach is necessary due to the fact that there are currently no reference measurement procedures available that can independently assign a reference value to viral reference materials for molecular in vitro diagnostic tests. Digital PCR (dPCR) is a technique that has the potential to be used for this purpose. In this paper, we investigate the ability of reverse transcriptase dPCR (RT-dPCR) to quantify HIV-1 genomic RNA without calibration. Criteria investigated included the performance of HIV-1 RNA extraction steps, choice of reverse transcription approach and selection of target gene with assays performed in both single and duplex format. We developed a protocol which was subsequently applied by two independent laboratories as part of an external quality assurance (EQA) scheme for HIV-1 genome detection. Our findings suggest that RT-dPCR could be used as reference measurement procedure to aid the value assignment of HIV-1 reference materials to support routine calibration of HIV-1 viral load testing by RT-qPCR. |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Publication date: | 01.05.2022 |
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| Year of publishing: | 2022 |
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| Number of pages: | str. [34]-40 |
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| Numbering: | Vol. 201 |
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| PID: | 20.500.12556/DiRROS-19309  |
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| UDC: | 577 |
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| ISSN on article: | 1046-2023 |
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| DOI: | 10.1016/j.ymeth.2021.03.006 |
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| COBISS.SI-ID: | 83727875 |
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| Note: | Soavtorji: Rainer MacDonald, Eloise J. Busby, Denise M. OʼSullivan, Mojca Milavec, Annabell Plauth, Martin Kammel, Heinz Zeichhardt, Hans-Peter Grunert, Andreas Kummrow, Jim F. Huggett;
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| Publication date in DiRROS: | 16.07.2024 |
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| Views: | 320 |
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| Downloads: | 168 |
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