Comparative evaluation of three JAK2V617F mutation detection methods

• Published in: American journal of clinical pathology Vol. 128; no. 5; pp. 865 - 874
• Main Authors: FRANTZ, Christine, SEKORA, Donna M, HENLEY, Donald C, HUANG, Chih-Kang, QIULU PAN, QUIGLEY, Neil B, GORMAN, Eric, HUBBARD, Roger A, MIRZA, Imran
• Format: Journal Article
• Language: English
• Published: Chicago, IL American Society of Clinical Pathologists 01.11.2007
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Alleles; Amplified Fragment Length Polymorphism Analysis - methods; Biological and medical sciences; Child; DNA Mutational Analysis - methods; Electrophoresis; Female; Humans; Investigative techniques, diagnostic techniques (general aspects); Janus Kinase 2 - blood; Janus Kinase 2 - genetics; Male; Medical sciences; Middle Aged; Mutation; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Polymorphism, Restriction Fragment Length - genetics; Predictive Value of Tests; Reagent Kits, Diagnostic; Reproducibility of Results; Evaluation; Polyacrylamide gel electrophoresis; Enzyme; Capillary electrophoresis; Gel electrophoresis; JAK2 protein tyrosine kinase; Transferases; Method; Polymerase chain reaction; Allele-specific PCR; Allele; Anatomic pathology; JAK2 mutation; Restriction fragment length polymorphism; AS-PCR; Acrylamide polymer; Genetics; Diagnosis; Mutation; Detection; Protein-tyrosine kinase; Comparative study; RFLP
• ISSN: 0002-9173; 1943-7722

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.