Samreen Falak (Avtor), Rainer Macdonald (Avtor), Eloise J. Busby (Avtor), Denise M. O'Sullivan (Avtor), Mojca Milavec (Avtor), Annabell Plauth (Avtor), Martin Kammel (Avtor), Heinz Zeichhardt (Avtor), Hans-Peter Grunert (Avtor), Andreas Kummrow (Avtor), Jim F. Huggett (Avtor)

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.

Ključne besede

imunski sistem;digitalni PCR;virusi;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: NIB - Nacionalni inštitut za biologijo
UDK: 577
COBISS: 83727875 Povezava se bo odprla v novem oknu
ISSN: 1046-2023
Št. ogledov: 43
Št. prenosov: 10
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarne ključne besede: imunski sistem;digitalni PCR;virusi;
Komentar vira: 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;
Strani: str. [34]-40
Zvezek: ǂVol. ǂ201
Čas izdaje: May 2022
DOI: 10.1016/j.ymeth.2021.03.006
ID: 24536857