Visual screening for JAK2V617F mutation by a disposable dipstick.
During the last 5 years, it was discovered that the JAK2V617F somatic mutation is present in virtually all patients with polycythemia vera and a large proportion of patients with essential thrombocythemia, primary myelofibrosis, and refractory anemia with ring sideroblasts and thrombocytosis. As a result, JAK2V617F was incorporated as a new clonal marker in the 2008 revision of the WHO diagnostic criteria. Current methods for JAK2 genotyping include direct sequencing, pyrosequencing, allele-specific PCR with electrophoresis, restriction fragment length polymorphism, real-time PCR, DNA-melting curve analysis, and denaturing HPLC. Some of these methods are labor intensive and time consuming, while the others require specialized costly equipment and reagents. We report a method for direct detection of the JAK2V617F allele by the naked eye using a dipstick test in a dry-reagent format. The method comprises a triprimer PCR combined with visual detection of the products within minutes by the dipstick test. Specialized instrumentation is not involved. The requirements for highly qualified technical personnel are minimized. Because the detection reagents exist in dry form on the dipstick, there is no need for multiple pipetting and incubation steps.
Konstantou
Jessica K
JK
Laboratory of Analytical Chemistry, Department of Chemistry, Athens University, 15771, Athens, Greece.
Iliadi
Alexandra C
AC
Ioannou
Penelope C
PC
Christopoulos
Theodore K
TK
Anagnostopoulos
Nikolaos I
NI
Kanavakis
Emmanuel
E
Traeger-Synodinos
Jan
J
D000483:Alleles; D017931:DNA Primers / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D020125:Mutation, Missense; D011087:Polycythemia Vera / Q000175:diagnosis / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
D017931:DNA Primers; D053614:Janus Kinase 2