| 用裂解法获得有核细胞进行实时定量RT-PCR的可行性研究 |
| |
| 引用本文： | 李玲娣,秦亚溱,李金兰,刘艳荣. 用裂解法获得有核细胞进行实时定量RT-PCR的可行性研究[J]. 中国实验血液学杂志, 2011, 19(2): 477-479 |
| |
| 作者姓名： | 李玲娣 秦亚溱 李金兰 刘艳荣 |
| |
| 作者单位： | 北京大学人民医院,血液病研究所,北京,100044 |
| |
| 摘 要： | 本研究比较裂解法获得的有核细胞与分层法获得的单个核细胞进行实时定量RT-PCR(RQ-PCR)的结果之间是否存在差异性.14例白血病患者(AML-M2 7例,APL 1例,AML M4 Eo 1例,AML M4 1例,AML M6 1例,CML3例)的骨髓标本,分别分成两等份,并分别采用分层法和裂解法获得细胞提取RNA进行实时定量RT-PCR,均检测ABL内参基因,并分别检测3份BCR-ABL(P210)、6份AML-ETO、1份CBFβ-MHY11、5份WT1及6份PRAME mRNA水平.结果显示:①28份样本ABL的拷贝数均>3×104,14对标本之间的内参基因ABL拷贝数没有显著性差异(p>0.05),因此两种方法均能保证RQ-PCR足够的敏感性.②14对标本检测的各目的基因mRNA水平亦没有显著性差异(p>0.05),说明两种方法最终得到的mRNA表达水平基本一致,对RQ-PCR结果没有影响.结论:采用裂解法获得有核细胞提取RNA是一种可行的方法,适用于同时提取大量标本RNA并进行RQ-PCR测定白血病特异基因的临床常规检测中.
|
| 关 键 词： | 裂解法 分层法 实时定量RT-PCR |
Evaluating the Feasibility of Nucleated Cells Isolated by Lysis Method for Real-time Quantitative RT-PCR |
| |
| LI Ling-Di,QIN Ya-Zhen,LI An-Lan,LIU Yan-Rong. Evaluating the Feasibility of Nucleated Cells Isolated by Lysis Method for Real-time Quantitative RT-PCR[J]. Journal of experimental hematology, 2011, 19(2): 477-479 |
| |
| Authors: | LI Ling-Di QIN Ya-Zhen LI An-Lan LIU Yan-Rong |
| |
| Affiliation: | LI Ling-Di,QIN Ya-Zhen,LI Jin-Lan,LIU Yan-Rong Institute of Hematology,Peking University People Hospital,Beijing 100044,China |
| |
| Abstract: | This study was aimed to investigate whether differance exists between real time RT-PCR results of nucleated cells isolated by lysis method and mononuclear cells isolated by gradient concentration method.14 bone marrow samples from leukemia patients(7 samples of AML-M2,1 of AML-M4,1 of AML-M4EO,1 of AML-M6,1 of APL and 3 of CML) were collected.Each sample was divided into 2 parts,and was used to isolate mononuclear cells by Ficoll-Hypaque gradient centrifugation,and other was used to isolate nuclear cells by... |
| |
| Keywords: | |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
