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  • Title: Rapid detection of JAK2 V617F mutation using high-resolution melting analysis with LightScanner platform.
    Author: Qian J, Lin J, Yao DM, Chen Q, Xiao GF, Ji RB, Li Y, Yang J, Qian Z.
    Journal: Clin Chim Acta; 2010 Dec 14; 411(23-24):2097-100. PubMed ID: 20728437.
    Abstract:
    BACKGROUND: Detection of the JAK2 mutation has recently been included under the essential diagnostic criteria for myeloproliferative neoplasm (MPN). High-resolution melt (HRM) curve analysis, a nongel-based, automated system, is introduced as a means of mutation scanning without the requirement of any post-PCR handling. METHODS: We studied the sensitivity and reproducibility of LightScanner™ platform in the detection of JAK2 V617F mutation and the availability for diagnostic use in MPN. RESULTS: The reproducible sensitivity of HRM analysis with LightScanner™ platform was 5% for the detection of JAK2 V617F mutation. In the test of blind screening of 105 samples (48 Ph- MPN and 57 Ph+ chronic myeloid leukemia), the identical judgement was interpreted by two blinded investigators. HRM analysis of all cases was fully concordant with the results of PCR-RFLP and direct sequencing. CONCLUSIONS: The HRM method developed here is an extremely sensitive, accurate and reliable technique and allows high-throughput, fast pre-screening to select for sequencing only those specimens that most likely contain mutant JAK2 V617F allele(s).
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