Comparative evaluation of three JAK2V617F mutation detection methods.
The correlation of JAK2V617F with a proportion of chronic myeloproliferative disorders has generated numerous studies focused on the development of molecular-based assays for JAK2V617F detection. The current parallel study comparatively evaluated 3 JAK2V617F molecular detection methods. Genomic DNA from blood or bone marrow was assayed by 3 laboratories using allele-specific polymerase chain reaction (AS-PCR) or kit-based restriction fragment length polymorphism methods, which used polyacrylamide gel or capillary electrophoresis analysis. In addition, samples were sequenced in 2 of the laboratories. Results found 100% concordance among the 3 methods, with analytic sensitivities of 5% for both kit methods and 0.01% for AS-PCR. The kitbased assays detect JAK2V617F with equal sensitivity regardless of analysis method, and, despite greater sensitivity of AS-PCR, all 3 methods yielded 100% concordant results for this 36-sample set. Consistent with other reports, direct sequencing was insufficiently sensitive to serve as an initial diagnostic tool for JAK2V617F detection.
Frantz
Christine
C
Department of Laboratory Medicine and Pathology, University of Alberta Hospital, Edmonton, Canada.
Sekora
Donna M
DM
Henley
Donald C
DC
Huang
Chih-Kang
CK
Pan
Qiulu
Q
Quigley
Neil B
NB
Gorman
Eric
E
Hubbard
Roger A
RA
Mirza
Imran
I
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D054458:Amplified Fragment Length Polymorphism Analysis / Q000379:methods*; D002648:Child; D004252:DNA Mutational Analysis / Q000379:methods; D004586:Electrophoresis; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000097:blood / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results
D011933:Reagent Kits, Diagnostic; C507924:JAK2 protein, human; D053614:Janus Kinase 2