| Naslov: | An assessment of the reproducibility of reverse transcription digital PCR quantification of HIV-1 |
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| Avtorji: | ID Falak, Samreen (Avtor)
ID Macdonald, Rainer (Avtor)
ID Busby, Eloise J. (Avtor)
ID O'Sullivan, Denise M. (Avtor)
ID Milavec, Mojca (Avtor)
ID Plauth, Annabell (Avtor)
ID Kammel, Martin (Avtor)
ID Zeichhardt, Heinz (Avtor)
ID Grunert, Hans-Peter (Avtor)
ID Kummrow, Andreas (Avtor)
ID Huggett, Jim F. (Avtor) |
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| Datoteke: |  URL - Izvorni URL, za dostop obiščite https://doi.org/10.1016/j.ymeth.2021.03.006
 PDF - Predstavitvena datoteka, prenos (869,23 KB)
MD5: 59CF6A5B70A39431A8C01A99430F6BA8
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| Jezik: | Angleški jezik |
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| Tipologija: | 1.01 - Izvirni znanstveni članek |
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| Organizacija: |  NIB - Nacionalni inštitut za biologijo
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| Povzetek: | 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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| Status publikacije: | Objavljeno |
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| Verzija publikacije: | Objavljena publikacija |
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| Datum objave: | 01.05.2022 |
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| Leto izida: | 2022 |
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| Št. strani: | str. [34]-40 |
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| Številčenje: | Vol. 201 |
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| PID: | 20.500.12556/DiRROS-19309  |
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| UDK: | 577 |
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| ISSN pri članku: | 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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| Opomba: | 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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| Datum objave v DiRROS: | 16.07.2024 |
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| Število ogledov: | 325 |
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| Število prenosov: | 169 |
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| Metapodatki: |    |
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