Accurate quantitation of JAK 2 V617F allele burden by array‐based digital PCR

Summary Introduction The JAK 2 V617F mutation is highly prevalent in patients with myeloproliferative neoplasms ( MPN ). Several studies have shown that allele burden correlates with hematologic characteristics and clinical end‐points in patients with MPN . Allele‐specific real‐time quantitative polymerase chain reaction ( RQ ‐ PCR ) is probably the most commonly used technique for detection and quantitation of the JAK 2 V617F mutation. Alternatively, digital PCR is an emerging technology for absolute DNA quantitation, which is based on dilution and high‐grade partitioning of a sample. Methods We compared array‐based digital PCR using the QuantStudio ™ 3D Digital PCR System platform with a RQ ‐ PCR assay, CE ‐registered for in vitro diagnostic use, regarding JAK 2 V617F allele quantitation. This study included 30 samples positive for the JAK 2 V617F mutation and additionally 13 samples without the mutation. Results JAK 2 V617F allele burden of samples ranged between well below 1% and more than 90%. Linear regression analysis showed a high correlation between the results obtained from the two techniques ( r 2  = 0.9983). Thirteen samples tested negatively for the JAK 2 V617F mutation with RQ ‐ PCR were also found negative with digital PCR . Conclusion We conclude that array‐based digital PCR is an appropriate method for the quantitation of JAK 2 V617F allele burden demonstrating highest correlation with allele‐specific RQ ‐ PCR .