定量检测bcr—abl（P210）转录本水平多中心比对研究  

A multicenter comparison study on the quantitative detection of bcr-abl （e210） transcript levels in China

作　　者：秦亚溱[1] 程辉[2] 岑建农[3] 耿素霞[4] 李庆华[5] 李小青 林振兴[7] 马道新[8] 乔纯[9] 王云贵[10] 李金兰[1] 李玲娣[1] 黄晓军[1] 

机构地区：[1]北京大学人民医院,北京大学血液病研究所,100044 [2]第二军医大学附属长海医院血液科 [3]苏州大学附属第一医院血液病研究所 [4]广东省人民医院血液科 [5]中国医学科学院、北京协和医学院血液学研究所、血液病医院 [6]武汉市协和医院血液科 [7]福建医科大学附属协和医院血液病研究所 [8]山东大学齐鲁医院血液科 [9]南京医科大学第一附属医院 江苏省人民医院血液科 [10]浙江大学附属第一医院血液科

出　　处：《中华血液学杂志》2013年第2期104-108,共5页Chinese Journal of Hematology

基　　金：北京市科技计划项目（Z111107067311070）

摘　　要：目的探讨国内不同医院bcr—abl（P210）转录本水平定量检测值的可比性。方法2010年4月至2011年8月，10家医院进行了4次样本派发以比对bcr—abl（P210）转录本水平定量检测值。由北京大学人民医院统一制备4个梯度（10倍系列稀释）的bcr—abl（P210）转录本水平细胞样本，加入TRIzol中，每个梯度每家医院各派发3份平行样品，每份样品细胞数为8×10^6，各单位分别按照自身实验方案采用实时定量PCR技术检测bcr—abl转录本水平。应用回归方程计算各单位转换系数（CF）。结果每次比对，各医院检测结果之间均存在差异。除个别医院外，各医院检测结果总体一致并与细胞稀释度一致。各家医院的CF均存在波动，其中3家医院比较稳定，4次比对CF范围分别为2．8～5．2、1．2—2．8和2．2～6．8；两家医院的CF变化较大，范围分别为0．76～7．0和2．1～18．7；三家医院各有1～2次检测结果明显偏离实际水平而无法计算出CF，能够算出的CF范围分别为1．9～19．2、3．6～7．6和0．18～14．7；一家医院因为中途更换主要试剂只有2次比对的CF（3．3和5．0）。结论通过比对获得CF能够实现不同医院bcr—abl转录本水平检测值的可比性。检测体系稳定可靠是获得准确CF的前提。Objective To investigate the comparability of bcr-abl （19210） transcript levels detected in different hospitals. Methods Ten hospitals in China took part in the four times of sample exchange and comparisons fi＇om April, 2010 to August, 2011. The exchange samples were prepared by Peking University People＇ s Hospital. Firstly, the BCR-ABL （P210） （ ＋ ） cells from a newly diagnosed chronic myeloid leuke- mia patient were 10-fold serially diluted by BCR-ABL （P210） （ - ） cells and they covered 4 magnitudes. Then, TRIzol reagents were thoroughly mixed with cells in each tube. Every 12 samples （three samples per magnitude） were sent to the other 9 hospitals. The cell number of each sample was 8 × 10^6. The detection of bcr-abl transcript levels by real-time quantitative PCR were performed in every hospital according to their own protocols. Conversion factors （CF） were calculated using regression equation. Results Differences in bcr- abl transcript levels did exist among results of 10 hospitals in each comparison. In general, the results of the most of hospitals were in line with the dilutions of cells. CF of every hospital fluctuated. Three hospitals had relatively stable CF, and their ranges were 2.8-5.2, 1.2-2.8 and 2.2-6.8, respectively; two hospitals had unstable CF with ranges 0.76 - 7.0 and 2.1 - 18.7 ; three hospitals couldn＇ t be calculated CF one or two times because of the significant deviation of the results from the actually bcr-abl transcript levels, and their ranges of CF which could be calculated were 1.9 - 19.2,3.6 - 7.6 and O. 18 - 14.7 ; One hospital only had two CF （3.3 and 5.0） because of the replacement of an important reagent during the period of comparisons. Conclusions Comparability of bcr-abl （P210） transcript levels between different hospitals could be achieved through CF which acquired by sample exchange and comparison. The stable and reliable detection system is the premise to acquire correct CF.

关 键 词：融合基因 bcr—abl(P210) 聚合酶链反应 实时定量 转换系数 

分 类 号：R733.72[医药卫生—肿瘤]