High resolution melting analysis for rapid and effective detection of JAK2 V617F mutation in clinical samples

Objectives To explore the application of real time PCR technique and high resolution melting(HRM) analysis in the detection of JAK2 V617F mutation for clinical samples.Methods Two different cell lines(HEL and HL60) were used to validate the capacity of HRM method for detection of JAK2 V617F mutation.The DNA samples from 63 patients clinically suspected with myeloproliferative neoplasm(MPN) were detected by HRM method.The results of HRM analysis were compared with DNA direct sequencing.Results Following PCR amplification,as low as 5% of JAK2 V617F mutation was successfully detected in the suspected MPN patients with MPN using the HRM analysis.High repetition was demonstrated in HRM analysis.The comparability of 100% with the direct sequencing was shown.Conclusion JAK2 V617F mutation could be detected with HRM analysis by performing real time PCR in the same tube on LightCycler 480 analyzer.The method was proven to be sensitive and specific for clinical samples.