Multiplex sequence-specific PCR detection for simultaneous screening of 5 types of JAK2 mutations in myeloproliferative diseases
10.3760/cma.j.issn.1009-9158.2009.01.007
- VernacularTitle:多重位点特异性PCR检测骨髓增殖性疾病五种JAK2基因突变
- Author:
Hongxing LIU
;
Chunrong TONG
;
Peng CAI
;
Guirong TANG
;
Xian ZHANG
;
Wen TENG
;
He WANG
;
Ying ZHANG
;
Ping ZHU
- Publication Type:Journal Article
- Keywords:
Myeloprolfferative disorders;
Janus kinase 2;
Mutation;
Polymerase chain reaction
- From:
Chinese Journal of Laboratory Medicine
2009;32(1):30-34
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.
