RUNX1-RUNX1T1融合转录本及WT1转录本水平检测的室间比对研究  

An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels

作　　者：秦亚溱[1] 朱立文 林樉[3] 耿素霞 刘生伟[5] 程辉 邬成业[7] 肖敏 李小青 胡瑞萍[10] 王丽丽[11] 刘海燕[12] 马道新[13] 关涛[14] 叶远馨[15] 牛挺[16] 岑建农[17] 卢利莎 孙黎 杨同华[20] 王云贵[21] 李涛[22] 王玥[23] 李庆华 赵晓甦[1] 李玲娣[1] 陈文敏 龙玲玉 黄晓军[1] Qin Yazhen;Zhu Liwen;Lin Shuang;Geng Suxia;Liu Shengwei;Cheng Hui;Wu Chengye;Xiao Min;Li Xiaoqing;Hu Ruiping;Wang Lili;Liu Haiyan;Ma Daoxin;Guan Tao;Ye Yuanxin;Niu Ting;Cen Jiannong;Lu Lisha;Sun Li;Yang Tonghua;Wang Yungui;Li Tao;Wang Yue;Li Qinghua;Zhao Xiaosu;Li Lingdi;Chen Wenmin;Long Lingyu;Huang Xiaojun(Peking University People’s Hospital,Peking University Institute of Hematology,National Clinical Research Center for Hematologic Disease,Beijing 100044,China;Beijing Hightrust Diagnostics Co.,Ltd,Beijing 100176,China;Department of Hematology,The First Affiliated Hospital of Dalian Medical University,Dalian 116011,China;Department of Hematology,Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;Harbin Institute of Hematology and Oncology,Harbin 150010,China;Changhai Hospital,The Second Military Medical University,Shanghai 200433,China;Institute of Hematology,Henan Provincial People’s Hospital,Zhengzhou 450003,China;Department of Hematology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030;Center for Stem Cell Research and Application,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022;Department of Hematology,Bethune First Affiliated Hospital of Jilin University,Changchun 130021;Department of Hematology,Chinese PLA General Hospital,Beijing 100853,China;Department of Hematology,Affiliated Hospital of Nantong University,Nantong 226001,China;Department of Hematology,Qilu Hospital of Shandong University,Jinan 250012,China;Department of Hematology,Shanxi Provincial Cancer Hospital,Taiyuan 030000,China;Department of Laboratory Medicine,West China Hospital of Sichuan University,Chengdu 610041,China;Department of Hematology,West China Hospital of Sichuan University,Chengdu 610041,China;The First Affiliated Hospital of Soochow University,Jiangsu Institute of Hematology,National Clinical Research Center for Hematologic Disease,Suzhou 215006,China;Tianjin Sino-us Diagnostics C

机构地区：[1]北京大学人民医院,北京大学血液病研究所,国家血液系统疾病临床医学研究中心,100044 [2]北京海思特医学检验实验室有限公司,100176 [3]大连医科大学附属第一医院血液科,116011 [4]广东省人民医院(广东省医学科学院)血液科,广州510080 [5]哈尔滨血液病肿瘤研究所,150010 [6]海军军医大学附属长海医院血液内科,上海200433 [7]河南省人民医院血液病研究所,郑州450003 [8]华中科技大学同济医学院附属同济医院血液科,武汉430030 [9]华中科技大学同济医学院附属协和医院干细胞中心,武汉430022 [10]吉林大学第一医院血液科,长春130021 [11]解放军总医院第一医学中心血液病科,北京100853 [12]南通大学附属医院血液科,226001 [13]山东大学齐鲁医院血液科,济南250012 [14]山西省肿瘤医院血液科,太原030000 [15]四川大学华西医院实验医学科,成都610041 [16]四川大学华西医院血液科,成都610041 [17]苏州大学附属第一医院,江苏省血液研究所,国家血液系统疾病临床医学研究中心,215006 [18]天津协和华美医学诊断技术有限公司,301617 [19]武汉康圣达医学检验所有限公司,430075 [20]云南省第一人民医院血液科,昆明650034 [21]浙江大学医学院附属第一医院血液科,杭州310003 [22]郑州大学第一附属医院血液科,450052 [23]中国医科大学附属第一医院,沈阳110001 [24]中国医学科学院血液病医院(血液学研究所),国家血液系统疾病临床医学研究中心,天津300020

出　　处：《中华血液学杂志》2019年第11期889-894,共6页Chinese Journal of Hematology

基　　金：国家自然科学基金(81870125)。

摘　　要：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。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.Results①RUNX1-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≥0.98.③The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of W

关 键 词：融合蛋白质类 RUNX1-RUNX1T1 WT1 实时聚合酶链反应 室间比对 

分 类 号：R4[医药卫生—临床医学]