Detection of the JAK2 V617F mutation in chronic myeloproliferative disorders by high resolution melting curve analysis

Aim To establish a rapid and simple diagnostic assay for detection of acquired JAK2V617F mutation of chronic myeloproliferative disorders(CMPDs) using high resolution melting curve analysis(HRM).Methods HRM technology was used to detect JAK2V617F mutation in 36 CMPDs samples detected by allele-specific-PCR assay(AS-PCR) previously,and compared with AS-PCR.The mutations in all samples were validated by direct sequencing.Results The segments of JAK2 gene were successfully amplified,and high-resolution melting curves for all samples were generated,and wild-type samples were definitely and reproducibly discriminated from JAK2V617F-positive samples.The sensitivity and specificity of HRM achieved 100%.When compared to AS-PCR,the coincidence rate of JAK2V617F mutations detected by two assays was 97%(35/36),and only one JAK2V617F-positive sample didn't match,which actually existed a JAK2V617F mutation verified by sequencing.Conclusion The HRM analysis is a rapid,simple and sensitive method for detection of JAK2V617F mutation.