Validity test study of JAK2 V617F and allele burden quantification in the diagnosis of myeloproliferative diseases
Validity test study of JAK2 V617F and allele burden quantification in the diagnosis of myeloproliferative diseases
复制标题
DOI：
10.1007/s00277-008-0512-x
复制
发表时间：
2008-09-01
影响因子：
3.5
通讯作者：
Martinez-Lopez, Joaquin
中科院分区：
文献类型：
--
作者：
Rapado, Inmaculada;Albizua, Enriqueta;Martinez-Lopez, Joaquin
近年来已有几种检测JAK2 V617F突变的灵敏方法问世，其中大多数是基于实时聚合酶链式反应(Real Time PCR)。然而，只有他们中的一些人进行了诊断有效性的研究。本研究比较了三种基于实时定量聚合酶链式反应检测JAK2 V617F突变的方法：一种是基于杂交探针(HP)和多肽核酸探针(PNA)的方法；另一种是采用等位基因特异的寡核苷酸引物进行JAK2 V617F的定量。149例健康人，其中原发性血小板增多症(ET)61例，真性红细胞增多症(PV)32例，继发性血小板增多症38例，继发性红细胞增多症35例。JAK2 617 HP聚合酶链式反应检测PV的敏感性(Se)为88%，特异性(Sp)为100%。Et中Se为57%，Sp为100%。JAK2617PNA在PV中的Se和Sp分别为94%和97.8%。Et中Se占70%，Sp占95.7%。在JAK2V671F等位基因特异性寡核苷酸(ASO)定量聚合酶链式反应(QPCR)中，通过接收工作特性(ROC)曲线确定了1%的截断点。在PV中，Se为93.8%，Sp为98.5%。Et中Se占80%，Sp占95.9%。2%的健康受试者JAK2 617 PNA聚合酶链式反应阳性，2%的JAK2 617反义寡核苷酸聚合酶链式反应阳性。通过克隆和测序，检测了健康受试者JAK2 V617F突变。JAK2617HP对红细胞增多症和血小板增多症均有较好的鉴别诊断价值。当JAK2 V617F等位基因负荷较低时，应进行JAK2 617 ASO qPCR。同时检测JAK2V617F和PRV-1的过度表达并不能提高JAK2V617F试验对MPD的诊断价值。
Several sensitive methods for the detection of JAK2 V617F mutation have been published recently, most of them based on Real Time polymerase chain reaction (PCR). However, only some of them have performed studies of diagnostic validity. This study compares three methods based on Real Time PCR to detect JAK2 V617F mutation: two based on hybridization probes (HP) and peptide nucleic acid probe (PNA) and a third employing allele specific oligonucleotide primers for JAK2 V617F quantification. One hundred forty-nine healthy subjects, 61 essential thrombocythemia (ET), 32 polycythemia vera (PV), 38 secondary thrombocytoses, and 35 secondary erythrocytoses were included. Validity test study for JAK2 617 HP PCR in PV Sensitivity (Se) was 88% and in Specificity (Sp), 100%. In ET, Se was 57% and Sp, 100%. For JAK2 617 PNA PCR in PV, Se was 94% and Sp, 97.8%. In ET, Se was 70% and Sp, 95.7%. In JAK2 V671F allelo-specific-oligonucleotide (ASO) quantitative PCR (qPCR), cutoff point of 1% was established by receiving operating characteristic (ROC) curves. In PV, Se was 93.8% and Sp, 98.5%. In ET, Se was 80% and Sp, 95.9%. Two percent of the healthy subjects were positive by JAK2 617 PNA PCR and 2% by JAK2 617 ASO qPCR. JAK2 V617F mutation was detected in healthy subjects by cloning and sequencing. JAK2 617 HP is an adequate test in differential diagnosis for both erythrocytosis and thrombocytosis. When JAK2 V617F allele burden is low, JAK2 617 ASO qPCR should be performed. Simultaneous determination of JAK2 V617F and PRV-1 overexpression does not improve the diagnostic value of JAK2 V617F tests in MPD.