Design and application of a novel PNA probe for the detection at single cell level of JAK2V617F mutation in Myeloproliferative Neoplasms.
Design and application of a novel PNA probe for the detection at single cell level of JAK2V617F mutation in Myeloproliferative Neoplasms.
复制标题
DOI：
10.1186/1471-2407-13-348
复制
发表时间：
2013-07-18
期刊：
影响因子：
3.8
通讯作者：
Cilloni D
中科院分区：
文献类型：
--
作者：
Bracco E;Rosso V;Serra A;Carnuccio F;Gaidano V;Nicoli P;Musto P;Saglio G;Frassoni F;Cilloni D
JAK2基因(V617F)的突变(S)在相当大比例的费城阴性骨髓增殖性肿瘤患者中已被描述，其检测现在是诊断算法的基石。我们开发了一种基于多肽核酸(PNA)技术和免疫荧光显微镜(PNA-FISH)相结合的新方法，用于在单细胞水平上特异性检测JAK2突变，从而提高了诊断分辨率和克隆患病率的研究。结果发现，真性红细胞增多症患者CD34+细胞突变百分率为40%~100%，原发性血小板增多症患者CD34+细胞突变百分率为15%~80%，原发性骨髓纤维化患者CD34+细胞突变百分率为25%~100%。这种方法可以在单细胞水平上以高度特异性区分NPM患者中携带突变JAK2或野生型JAK2的CD34+祖细胞。这种方法可以识别多个基因异常，这对于了解任何类型癌症的病理生理学和进化都是至关重要的。
Mutation(s) of the JAK2 gene (V617F) has been described in a significant proportion of Philadelphia negative Myeloproliferative Neoplasms (MPN) patients and its detection is now a cornerstone in the diagnostic algorithm. We developed a novel assay based on peptide nucleic acid (PNA) technology coupled to immuno-fluorescence microscopy (PNA-FISH) for the specific detection at a single cell level of JAK2-mutation thus improving both the diagnostic resolution and the study of clonal prevalence. Using this assay we found a percentage of mutated CD34+ cells ranging from 40% to 100% in Polycythemia Vera patients, from 15% to 80% in Essential Thrombocythemia and from 25% to 100% in Primary Myelofibrosis. This method allows to distinguish, with a high degree of specificity, at single cell level, between CD34+ progenitor stem cells harbouring the mutated or the wild type form of JAK2 in NPM patients. This method allows to identify multiple gene abnormalities which will be of paramount relevance to understand the pathophysiology and the evolution of any type of cancer.
登录
查看更多内容
影响因子：
50.3
作者：
Levine, RL;Wadleigh, M;Gilliland, DG
通讯作者：
Gilliland, DG
影响因子：
20.3
作者：
Scott, Linda M.;Scott, Mike A.;Green, Anthony R.
通讯作者：
Green, Anthony R.
DOI：
10.1073/pnas.0601462103
发表时间：
2006-04-18
影响因子：
11.1
作者：
Jamieson, CHM;Gotlib, J;Weissman, IL
通讯作者：
Weissman, IL
影响因子：
64.8
作者：
EGHOLM, M;BUCHARDT, O;NIELSEN, PE
通讯作者：
NIELSEN, PE
影响因子：
168.9
作者：
Baxter, EJ;Scott, LM;Green, AR
通讯作者：
Green, AR