Evaluation of QIAcuity digital polymerase chain reaction platform for the quantitation of JC and BK polyomaviruses

Le résumé fourni par la source

Monitoring the replication of human polyomaviruses (HPyV) JCPyV and BKPyV, either due to infection or re-activation, is important in patients with different immunosuppressive treatments. We evaluated whether digital polymerase chain reaction (dPCR) would be a feasible alternative to quantitative PCR (qPCR) for detection and viral load determination of JCPyV and BKPyV, simultaneously. We adapted a laboratory-designed duplex qPCR assay to QIAcuity dPCR platform and evaluated the performance of dPCR using plasmid calibrators, clinical samples, and external quality control assessment (EQA) samples. Plasmids containing JCPyV or BKPyV genomes showed excellent linear correlation between reference values and dPCR results, with a >4 log(10) dynamic range and low variation between replicates. Of 17 EQA core samples, 16 were within the consensus values by dPCR, while qPCR results deviated by at least 1 SD in 11 samples. There was a high agreement in the detection of both viruses, as well as good linear correlation between qPCR and dPCR in clinical samples. However, systemic differences in copy number estimations, presumably due to calibration imprecision in qPCR, were observed. The QIAcuity dPCR showed excellent performance in both the detection and copy number determination of JCPyV and BKPyV. The ease of use and high accuracy of quantitation makes it an attractive alternative to qPCR.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.