Comparative study of the commercial tests detecting frequent mutations in JAK2 and MPL

Background. qPCR is widely used to detect JAK2 V617F and MPL W515L/K mutations in myeloproliferative neoplasms. This study evaluated the diagnostic performance of four commercially available Russian qPCR test systems. Methods. 45 positive DNA samples (allelic burden 2—45%) and 18 negative samples were analyzed. Target allele frequencies were established using ddPCR as mean from 5 replicates. Two laboratories tested the kits (manufactured by «Genetechnology» LLC, «Formula gena» LLC, «Inogene» LLC, «Syntol» LLC) as independent participants and they were supplied from a manufacturer’s blinded, centralized purchase. Results. Specificity was 100% for all kits. JAK2 V617F: Sensitivity was 96.7% for three out of four kits, but 86.7% for «Genetechnology». «Syntol» kit showed the largest proportional systematic error (β1=1.63), while «Inogene» kit showed none (β1=1.00). «Genetechnology» kit had the highest coefficient of variation (CV=59.8%); «Syntol» kit had the lowest one (CV=10.2%). For MPL W515L mutation detection « Genetechnology « kit displayed both proportional (β1=0.25) and constant (β0=38.34) systematic errors with CV=14.1%. «Inogene» kit showed only a proportional error β1=0.66 and CV=9.4%. For MPL W515K mutation «Genetechnology» kit had a constant error β0=38.83 and CV=13.5%. «Inogene» kit showed a proportional error β1=1.45 and CV=8.9%. Conclusion. For JAK2 V617F mutation detection, the «Formula gena» kit showed the lowest deviation from the target value. For MPL W515L/K mutations, the tests from «Inogene» and «Genetechnology» provided comparable results.