Purpose/Principle

JAK2 V617F mutation detection is primarily used for patients with suspected MPNs who are Philadelphia chromosome (BCR/ABL1) negative. Testing may be used to diagnose, to help direct the appropriate treatment, monitor response to therapy and in some cases to detect early relapse.

This assay uses two real-time allele-specific PCR reactions. DNA from each sample is tested in a Mutant Specific (MT) reaction and a Wildtype (WT) specific reaction. The WT-specific forward primer is designed to perfectly match the normal guanine containing sequence, and the mutant-specific forward primer is designed to match the thymine point mutation containing sequence. The reverse primer and probe are common to both reactions. Amplification in each allele-specific reaction is tracked by monitoring fluorescence generated from the probe. This probe is cleaved during amplification, releases the attached quencher, and emits a signal proportional to the amount of DNA produced during amplification.