Superiority of Droplet Digital PCR Over Real-Time Quantitative PCR for JAK2 V617F Allele Mutational Burden Assessment in Myeloproliferative Neoplasms: A Retrospective Study

Abstract

JAK2 V617F mutational status is an essential diagnostic index in myeloproliferative neoplasms (MPNs). Although widely used for detection of JAK2 V617F mutation in peripheral blood (PB), sensitive real-time quantitative PCR (qPCR) presents some methodological limitations. Recently, emerging alternative technologies, like digital droplet PCR (ddPCR), have been reported to overcome some of qPCR’s technical drawbacks. The purpose of this study was to compare the diagnostic utility of ddPCR to qPCR for JAK2 V617F detection and quantification in samples from MPNs patients. Sensitivity and specificity of qPCR and ddPCR in the detection of the mutation were assessed by using a calibrator panel of mutated DNA on 195 JAK2 positive MPN samples. Based on our results, ddPCR proved to be a suitable, precise, and sensitive method for detection and quantification of the JAK2 V617F mutation.

Digital Droplet PCR in Hematologic Malignancies: A New Useful Molecular Tool

“…In the 99 samples analyzed by both techniques, there was an optimal correlation between QT-PCR and ddPCR, with the latest technique showing half a log higher sensitivity than the former one (5 × 10 −4 vs. 1 × 10 −3 ). PV and MF presented a similar median mutation burden (40.45%), higher than that observed in ET (21.35%) [ 125 ], differences that we also confirmed by different grades of the spleen stiffness observed by ultrasonography [ 126 , 127 , 128 ]. Finally, a Korean group compared a ddPCR assay for JAK2 V617F mutation with the results from pyrosequencing, once again showing the superiority of ddPCR [ 129 ].…”

The Impact of JAK2 V617F, CALR, and MPL Mutations as Molecular Diagnostic Markers of Myeloproliferative Neoplasms in Kurdish Patients. A Single-center Experience

“…In PMF patients, there was a significant difference between positive and negative mutation individuals in terms of HCT% and Platelet Count. Examining these results with the results of other studies showed that the average of demographic and clinical variables in this study was higher than in other studies (34)(35)(36).…”

Molecular Detection Methods in HPV-Related Cancers

“…The excellent sensitivity and specificity of HPV detection using ddPCR in swabs without the need for invasive tissue biopsy have several potential applications for both diagnosis and disease surveillance. Furthermore, the ddPCR method is reported to be accurate, repeatable, reproducible ( 27 , 94 , 100 ), and cost-effective ( 23 , 25 , 26 , 90 ).…”

“…It has been demonstrated that ddPCR exhibits high sensitivity, accuracy, specificity, repeatability, and reproducibility compared to RT-PCR in quantifying HPV DNA ( 31 , 71 , 92 ), and therefore it can be used to test the self-sampled swabs. Since ddPCR method is reported to be accurate, repeatable, reproducible ( 27 , 94 , 100 ), and cost-effective ( 23 , 25 , 26 , 90 ), it is an ideal method for routine diagnostic testing.…”