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Clin Chim Acta. 2010 Dec 14;411(23-24):2097-100. doi: 10.1016/j.cca.2010.08.020. Epub 2010 Aug 19.

Rapid detection of JAK2 V617F mutation using high-resolution melting analysis with LightScanner platform.

Author information

1
Department of Hematology, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu 212002, PR China.

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 LightScannerâ„¢ platform in the detection of JAK2 V617F mutation and the availability for diagnostic use in MPN.

RESULTS:

The reproducible sensitivity of HRM analysis with LightScannerâ„¢ 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).

PMID:
20728437
DOI:
10.1016/j.cca.2010.08.020
[Indexed for MEDLINE]

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