JAK2 V617F mutation can be reliably detected in serum using droplet digital PCR

IntroductionDetection of the JAK2 V617F mutation is a key step in the diagnosis of myeloproliferative neoplasms (MPN). Sensitive real-time quantitative PCR (qPCR) detection on peripheral blood (PB) is the most widely used method. The main objective of this study was to determine whether serum, the most common material available in archival biobanks, is a good liquid biopsy for detecting and quantifying the JAK2 V617F mutation using droplet digital PCR (ddPCR). MethodsPaired PB and serum samples from 66 patients with MPN were used. Serum samples were frozen at -25 degrees C before analysis. DNA was extracted from 200L PB and 400L serum, and ddPCR analysis was performed. ResultsAmong the 47 patients with detectable mutation in their PB samples, the overall sensitivity for the detection of JAK2 mutation in serum was of 96% (45 of 47); V617F was detected in all cases where mutation load was above 1%. Our results showed very strong correlation between PB and serum (Spearman r: 0.989, P<.0001). Significantly higher allele burden was detected in serum compared to PB (Wilcoxon signed ranks test, Z=-5.672, P<.0001). ConclusionIn our study, JAK2 V617F mutation load as low as 1% was reliably detected in serum using ddPCR.