A multicenter comparison study on the quantitative detection of bcr-abl (P210) transcript levels in China

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8×106,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.To investigate the comparability of bcr-abl (P210) transcript levels detected in different hospitals.Ten hospitals in China took part in the four times of sample exchange and comparisons from 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 leukemia 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.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 0.18 - 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.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.北京市科技计划项目PubMed中文核心期刊要目总览(PKU)中国科技核心期刊(ISTIC)中国科学引文数据库(CSCD)02104-1083