WHO/BS/2016.2293 Collaborative study to evaluate the proposed WHO 1st International Reference Panel for Genomic JAK2 V617F

Overview

An international collaborative study assessed the suitability of a panel of genomic DNA materials as the proposed World Health Organization 1st International Reference Panel for Genomic JAK2 V617F, NIBSC code 16/120, for use in the standardization of myeloproliferative neoplasm-associated JAK2 V617F genomic-based research and diagnostics. The panel comprised seven freeze-dried genomic DNA materials at a range of JAK2 V617F nominal values (as a percentage of total JAK2); 100% JAK2 V617F (NIBSC material code 15/164), 90% (15/246), 30% (15/244), 10% (15/166), 1% (15/168), 0.1% (15/170) and 0% (15/172). Participants evaluated the materials using their routine diagnostic methods and against in-house controls (previously characterized patient samples and cell line-derived genomic DNAs), quality assessment materials or commercial materials. Where possible, results were reported quantitatively in order to assign a consensus value to each of the materials. Twenty nine laboratories in twenty countries performed thirty eight testing methods on the panel, of which thirty four reported quantitative data.

Conclusions from this study indicated that all seven materials were suitable for use as reference materials in the genomic diagnosis of JAK2 V617F using allele-specific quantitative PCR (ASQPCR), digital PCR, allelic discrimination-based endpoint PCR (including AS-PCR, Amplification Refractory Mutation System-PCR (ARMS-PCR) and Matrix Assisted Laser Desorption/IonizationTime of Flight (MALDI-TOF) mass spectrometric analysis) and next-generation sequencing (NGS), with proposed consensus values of 100% JAK2 V617F (15/164), 89.5% (15/246), 29.6% (15/244), 10.8% (15/166), 1.00% (15/168), 0.03% (15/170) and 0% (15/172), the median values of all (statistically appropriate) quantitative methods. All collaborative study participants agreed with the proposed consensus values and approved the panel as the proposed WHO 1st International Reference Panel for Genomic JAK2 V617F (NIBSC panel code 16/120).