Korean J Lab Med.  2009 Jun;29(3):243-248. 10.3343/kjlm.2009.29.3.243.

Usefulness of Real-time Semi-quantitative PCR, JAK2 MutaScreen (TM) Kit for JAK2 V617F Screening

• Chae H
• Lee JH
• Lim J
• Jung SW
• Kim M
• Kim Y
• Han K
• Cho BS
• Cho SG
• Lee JW
• Min WS

• KMID: 1096977
• DOI: http://doi.org/10.3343/kjlm.2009.29.3.243

Abstract

BACKGROUND: Real-time PCR for quantification of JAK2 V617F has recently been introduced and used to evaluate the importance of mutant allele burden in both diagnosis and disease progression in myeloproliferative diseases (MPDs). We evaluated the usefulness of JAK2 MutaScreen(TM) kit that uses a real-time semiquantitative PCR method and has been designed to screen JAK2 V617F mutant allele burden.
METHODS
Forty MPD patients were included in this study. We screened JAK2 V617F and determined the mutant allele burden using JAK2 MutaScreen(TM) kit. The mutant allele burden was estimated by six-scaled standards of JAK2 V617F mutant allele (2%, 5%, 12.5%, 31%, 50%, and 78%). For evaluation of test performance, an allele-specific PCR (AS-PCR) was carried out in all samples by using Seeplex JAK2 Genotyping kit. We assessed the clinical differences in distinct disease entities of MPDs according to JAK2 V617F mutant allele burden.
RESULTS
JAK2 V617F mutation was detected in 30 cases, including 10 of 11 cases (91%) of polycythemia vera (PV), 13 of 20 cases (65%) of essential thrombocythemia (ET), and 2 of 3 cases (67%) of chronic idiopathic myelofibrosis (CIMF). The concordance rate between the two tests was 95% (38/40). JAK2 V617F mutant allele burden was greater than 50% in 17 cases, and 10 of them (59%) were PV. In contrast, mutant allele burden was less than 50% in 13 cases and 11 of them (85%) were ET.
CONCLUSIONS
JAK2 MutaScreen(TM) kit that utilizes a real-time semi-quantitative PCR method is a useful tool for diagnosing MPDs precisely. It can be used to assess the grade of mutant allele burden as well as to screen JAK2 V617F simultaneously.

Keyword

JAK2 V617F; Myeloproliferative disease; Real-time PCR; Allele burden

MeSH Terms

Adult
Aged
*Alleles
Amino Acid Substitution
DNA Mutational Analysis
Disease Progression
Female
Humans
Janus Kinase 2/*genetics
Male
Middle Aged
Mutation
Myeloproliferative Disorders/*diagnosis/genetics
Polymerase Chain Reaction/*methods
Reagent Kits, Diagnostic

Figure

• 

  Fig. 1. Distribution of JAK2 V617F allele burden determined by JAK2 MutaScreen™ kit in 30 cases of JAK2 V617F positive MPDs. Abbreviations: PV, polycythemia vera; ET, essential thrombocythemia; CIMP, chronic idiopathic myelofibrosis; MPD-U, myeloproliferative disease, unclassifiable.

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