Tan Angela Y C, Westerman David A, Dobrovic Alexander
Department of Pathology, Division of Haematology and Medical Oncology, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
Am J Clin Pathol. 2007 Jun;127(6):977-81. doi: 10.1309/1U61JVXTLPPQ7YP1.
The point mutation 1849 (GT) V617F in the JAK2 gene occurs at high frequency in several chronic myeloproliferative diseases. Although a number of V617F mutation detection methods have been described, few are readily implemented in a diagnostic setting. We developed a simple and sensitive allelespecific competitive blocker polymerase chain reaction (ACB-PCR) assay to detect the V617F mutation. DNA was extracted from peripheral whole blood samples of 26 patients with chronic myeloproliferative disease. The ACB-PCR limit of detection was 1%. All positive samples detected by sequencing were detected by ACB-PCR. In 3 patients with essential thrombocythemia, the V617F mutation was readily detected by ACB-PCR but was near the detection limit of sequencing, confirming that ACB-PCR is more effective at detecting V617F when the mutant cell population is low. Detection of the monomorphic JAK2 V617F mutation using the ACB-PCR assay is easy to perform, rapid, sensitive, and cost-effective, which are key features of an ideal diagnostic method.
JAK2基因中的1849（GT）V617F点突变在几种慢性骨髓增殖性疾病中高频出现。虽然已经描述了多种V617F突变检测方法，但很少有方法能在诊断环境中轻易实施。我们开发了一种简单且灵敏的等位基因特异性竞争性阻断聚合酶链反应（ACB-PCR）检测法来检测V617F突变。从26例慢性骨髓增殖性疾病患者的外周全血样本中提取DNA。ACB-PCR的检测限为1%。所有通过测序检测到的阳性样本均能被ACB-PCR检测到。在3例原发性血小板增多症患者中，ACB-PCR能轻易检测到V617F突变，但该突变接近测序的检测限，这证实了当突变细胞群体比例较低时，ACB-PCR在检测V617F方面更有效。使用ACB-PCR检测法检测单态性JAK2 V617F突变操作简便、快速、灵敏且经济高效，这些都是理想诊断方法的关键特征。
