Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People's Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories' results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion: The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.
目的： 通过室间比对了解国内RUNX1-RUNX1T1融合转录本及WT1转录本检测现状和真实表现。 方法： 北京大学人民医院（简称PKUPH）制备比对样品，即用RUNX1-RUNX1T1（-）患者与RUNX1-RUNX1T1（+）患者新鲜骨髓/外周血有核细胞进行不同比例的稀释，制备出14种比对样本，每种样本各制备23份平行样本，加入TRIzol均质化后-70℃保存。各家中心采用RT-PCR技术同时检测各样本的RUNX1-RUNX1T1融合转录本及WT1转录本水平，统一以目的基因拷贝数/ABL拷贝数×100%的形式报告结果。通过Spearman相关分析计算各家中心与PKUPH检测结果之间的相关系数。 结果： ①RUNX1-RUNX1T1比对：9份为阳性、5份为阴性样本，参与的20家实验室的假阳性率为5%（5/100），假阴性率为0（0/180）。每份阳性样本各家检测值均不相同，9份阳性样本各家报告的结果中位值为0.060%~176.7%，共覆盖3.5个log的范围，各份样本最高与最低报告结果的比值为5.5~12.3（去除1份明显偏离的结果）。85%（17/20）的实验室与PKUPH结果之间的相关系数≥0.98。②WT1比对：14份样本各家报告结果均不相同，中位值为0.16%~67.6%，覆盖2.6个log的范围，各样本检测最高值与最低值的比值为5.3~13.7.62%（13/21）的实验室与PKUPH结果的相关系数≥0.98。③两个转录本每家与PKUPH报告结果的相对关系不一致，2家均低于、7家均高于PKUPH，另11家为1个高于另一个低于PKUPH。 结论： 同一样本各家中心报告的RUNX1-RUNX1T1及WT1转录本水平不同，大多数实验室与PKUPH报告的结果具有很高的一致性，实验室间不同转录本水平的相对关系不一定相同。.
Keywords: Fusion protein, RUNX1-RUNX1T1; Interlaboratory comparison; Real-time quantitative PCR; WT1.