实时定量PCR检测BCR-ABL（P210）转录本水平室内长期质控体系的建立及应用  

The establishment and application of internal quality control system for real-time quantitative PCR detection of BCR-ABL （P210）

作　　者：钟朝琴 何娜[1] 华明强 魏晓东[2] 马道新[1] 纪春岩[1] 

机构地区：[1]山东大学齐鲁医院血液科,济南250012 [2]山东大学附属济南市中心医院

出　　处：《中华血液学杂志》2016年第9期800-806,共7页Chinese Journal of Hematology

基　　金：国家自然科学基金（81470319）

摘　　要：目的制备BCR-ABL（P210）实时定量PCR（RQ-PCR）检测的室内质控物，建立RQ-PCR检测BCR-ABL（P210）转录本水平的室内质控方法。方法取K562细胞和HL-60细胞，制备RQ-PCR检测BCR-ABL（P210）转录本水平的高、低浓度质控物，2013年8月至2015年10月用RQ-PCR法与临床标本同步检}!测BCR-ABL（P210）184次，按试剂批号（批号20130303、20131212、20140411和20150327分别简称为R1、R2、R3、R4）统计分析标准曲线斜率、截距及相关系数，并按试剂批号和质控物批号（批号20130725、20140611分别简称为Q1、Q2）统计分析高、低浓度质控物检测结果，对斜率、截距及质控物检测结果采取Levey-Jennings质控图结合Westgard多规则质控方法进行质控判断。结果①标准曲线斜率、截距：R1检测53次，斜率、截距均未失控；R2检测37次，斜率失控6次，且第2-8次在x-s限值下侧，第12～37次在x值上侧，截距失控9次，且第1-8次在x＋s限值上侧，第12～37次出现在i值下侧；R3检测80次和R4检测14次，斜率、截距均未失控。②质控物结果：Q1批号质控物，R1检测49次朱失控，R2检测23次失控1次；Q2批号质控物，R2检测14次、R3检测72次及R4检测14次均未失控。结论利用K562细胞和HL-60细胞制备定量检测BCR-ABL（P210）转录本水平的高、低浓度室内质控物，制备方便，检测结果可靠、稳定，使用质控物检测结果结合标准曲线斜率、截距进行室内质控，可更有效地保证临床检测结果的准确性和稳定性。Objective To set internal quality control system of BCR-ABL （P210） transcript levels for real-time quantitative PCR （RQ-PCR）. Methods Using K562 cells and HL-60 cells, we prepared high- and low-level BCR-ABL internal quality control substance. The BCR-ABL （P210） transcript levels of internal quality control substance have been determined for 184 times together with clinical samples from August 2013 to October 2015. The slope rate, intercept and correlation coefficient of standard curve were calculated according to different reagent lots （lots number 20130303, 20131212, 20140411 and 20150327 are called RI ,R2 ,R3 and R4 for short respectively）, and the detection results of quality control substance were calculated according to different reagent lots and quality control substance lots （lots number 20130725, 20140611 are called Q1 ,Q2 for short respectively）. Then the results were analyzed by Levey-Jennings quality control chart combined with Westgard multi-rules theory. Results （1）We analyzed the slope rate and intercept of standard curve. Fifty-three times of the R1 reagent detection, 80 times of the R3 reagent detection and 14 times of the R4 reagent detection were all under control. For 37 times detection of R2 reagent, the slope rate was out of control for 6 times. It was lower than x-s for the 2-8 tests and upper the average for the 12-37 tests. The intercept was out of control for 9 times, upper the x＋s for the 1-8 tests and lower the average for the 12-37 tests. （2）According to the detection results of quality controlsubstance, for QI quality control substance, 49 tests by R1 reagent were under control, and 1 out of 23 tests by R2 reagent was out of control. For Q2 quality control substance, 14 tests by R2 reagent detection, 72 tests by R3 reagent detection and 14 tests by R4 reagent were all under control. Conclusion The preparation of high- and low-level quality control substance using K562 and HL-60 cells was convenient and the detection results were reliable and stable. The

关 键 词：实时聚合酶链反应 融合蛋白质类 BCR-ABL 室内质量控制 斜率 截距 

分 类 号：R733.72[医药卫生—肿瘤]