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The comparison of JAK2~(V617F)mutation detection methods in myeloproliferative disorders

LI Wei-da~1,LI Jian-yong~2,ZHANG Su-jiang~2,et al. (1.Department of Hematology,the Second People' s Hospital of Lianyungang,Jiangsu 222000,China; 2.Department of Hematology,The First Affiliated Hospital of Nanjing Medical Universit, Jiangsu People's Hospital,Nanjing 210029,China)  
Objective To investigate the better method to detect the occurrence of JAK2~(V617F) mutation in patients with myeloproliferative disorders(MPD).Methods Genomic DNA was extracted from peripheral blood cell samples of 16 case of chronic myelogenous leukemia(CML),22 cases of polycythaemia vera(PV),26 cases of essential thrombocythaemia(ET)and 12 cases of idiopathic myelofibrosis(IMF).Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and Allele-specific PCR(AS-PCR)were conducted to identify JAK2~(V617F) mutation individually.The sensitivity of two methods was compared successfully and the results were confirmed by sequence analysis consequently.Results The occurrence of JAK2~(V617F) mutation in PV patients detected by AS-PCR(72.7%)was higher than that of PCR-RFLP(50%).The occurrence of JAK2~(V6l7F) mutation in ET patients detected by AS-PCR(42.3%) was higher than that of PCR-RFLP(34.6%).All of the positive samples detected by PCR-RFLP were included in the positive samples of AS-PCR.The results of two methods were all coincided with sequence analysis. Conclusion AS-PCR is more sensitive,simple and convenient than PCR-RFLR.Consequently,it may have clinical significance in large scale detection of JAK2~(V617F) mutation.
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