Rapid detection of JAK2 V617F mutation using high-resolution melting analysis with LightScanner platform.
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).
Qian
Jun
J
Department of Hematology, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu 212002, PR China.
Lin
Jiang
J
Yao
Dong-Ming
DM
Chen
Qin
Q
Xiao
Gao-Fei
GF
Ji
Run-Bi
RB
Li
Yun
Y
Yang
Jing
J
Qian
Zhen
Z
D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000175:diagnosis / Q000235:genetics; D013997:Time Factors; D044366:Transition Temperature
D053614:Janus Kinase 2