Determining the JAK2 V617F mutation at Hue University of Medicine and Pharmacy Hospital

Abstract

Background: JAK2 V617F mutation is one of the major criteria in diagnosing MPNs. This study aimed to: (1) Establish a procedure and compare Allele specific-PCR and PCR-RFLP techniques in identifying JAK2 V617F mutation; (2) Primarily evaluate the prevalence of V617F mutation of JAK2 gene in patients with MPNs. Materials and methods: DNA extraction was isolated from peripheral blood of 31 patients with MPNs followed by clinicians and 10 healthy individuals. Determining V617F mutation of JAK2 gene by AS-PCR and PCR-RFLP techniques. Sanger sequencing randomly 15% of samples for comparison. Results: The optimal annealing temperature for AS-PCR is 55oC. Conditions of restriction digest in PCR-RFLP uses 300 ng of PCR product. All samples identified by AS-PCR and PCR-RFLP showed similar results. Conclusion: Established a procedure to identify the V617F mutation in JAK2 gene by allele-specific PCR and PCR-RFLP, in which the AS-PCR technique showed more advantages. The frequency of V617F mutation of JAK2 gene in patients was 61.3%. Key words: MPNs, V617F, JAK2, allele-specific PCR, PCR-RFLP