外周血样本的存储和运输对BCR-ABL(P210)转录本水平定量检测结果的影响  

The influence of peripheral blood sample storage and delivery on the quantitative detection result of BCR-ABL(P210)transcript level

作　　者：华明强 何娜[1] 钟朝琴 杨新雨 刘金婷 王锐卿 韩凤姣 张琛 马道新[1] Hua Mingqiang;He Na;Zhong Chaoqin;Yang Xinyu;Liu Jinting;Wang Ruiqing;Han Fengjiao;Zhang Chen;Ma Daoxin(Department of Hematology,Qilu Hospital,Cheeloo College of Medicine,Shandong University,Jinan,Shandong,250012,China;Department of Emergency Medicine,Qilu Hospital,Cheeloo College of Medicine,Shandong University,Jinan,Shandong,250012,China;Department of Hematology,Yantai Mountain Hospital,Yantai,Shandong 264000,China)

机构地区：[1]山东大学齐鲁医院血液科,济南250012 [2]山东大学齐鲁医院急诊科,济南250012 [3]山东省烟台山医院血液科,烟台264000

出　　处：《中华血液学杂志》2021年第3期224-229,共6页Chinese Journal of Hematology

摘　　要：目的探索慢性髓性白血病(CML)患者外周血样本的存储和运输对实时定量PCR(RQ-PCR)检测BCR-ABL(P210)转录本水平的影响。方法采集84例CML患者外周血样本,根据储存时间(0、6、12、24、48和72 h)、温度[室温(18~24℃)和低温(2~8℃)]以及振荡条件(振荡时长3、6和12 h)设置不同分组,RQ-PCR法检测不同条件下外周血样本BCR-ABL(P210)转录本水平。本研究将BCR-ABL拷贝数、ABL拷贝数和BCR-ABL(P210)转录本水平等原始数据进行对数转化(log10N)。结果①琼脂糖凝胶电泳结果显示:室温条件下,储存48、72 h的样本RNA出现显著降解;②储存温度方面,总体样本中,室温储存48、72 h的样本BCR-ABL拷贝数检测水平显著低于低温储存的样本(P<0.05);然而BCR-ABL(P210)转录本水平在低温储存与室温储存条件下检测结果差异无统计学意义(P>0.05);③储存时间方面,与基线的3.35±1.60相比,总体样本的BCR-ABL拷贝数检测结果仅在室温储存储存下48 h(2.93±1.59)和72 h(2.79±1.42)显著下降(P<0.05);与基线的5.47±0.35相比,总体样本中ABL拷贝数检测结果在48 h(低温5.29±0.30,室温5.06±0.38)和72 h(低温5.11±0.54,室温4.64±0.79)均显著下降(P<0.05)。但是,与基线的-0.56±1.51相比,无论低温或室温条件下,不同储存时间(6、12、24、48和72 h)的样本BCR-ABL(P210)转录本水平检测结果无显著改变(P>0.05);④振荡方面,与基线的-0.60±1.37相比,振荡3、6、12 h的样本BCR-ABL(P210)转录本水平检测结果无显著改变(P>0.05)。结论样本储存时间、储存温度及运输振荡因素可以影响BCR-ABL、ABL拷贝数的检测结果,但对BCR-ABL(P210)转录本水平定量检测结果无显著影响。Objective To explore the influence of storage and delivery conditions of the peripheral blood samples from patients with chronic myeloid leukemia(CML)on the real-time quantitative PCR(RQ-PCR)detection of the BCR-ABL(P210)transcript levels.Methods The peripheral blood samples of 84 CML patients were collected.The same sample was divided into different groups according to storage time(0,6,12,24,48,and 72 h),temperature(room temperature,18-24℃;low temperature,2-8℃),and vibration conditions(3,6,and 12 h).RQ-PCR was used to detect BCR-ABL(P210)transcript levels of the different groups.This study logarithmically transformed(log10N)the original data[BCR-ABL copy number,ABL copy number,and BCR-ABL(P210)transcript levels].Results①Agarose gel electrophoresis showed significant RNA degradation of samples after storage for 48 and 72 h at room temperature.②Among the overall samples,the BCR-ABL copy number of the samples stored at room temperature for 48 and 72 h was significantly lower than that of the samples stored at low temperature(P<0.05).However,the BCR-ABL(P210)transcript levels had no significant difference between samples stored at low temperature and room temperature.③No significant changes were noted in the BCR-ABL(P210)transcript levels at different storage times(6,12,24,48,and 72 h)regardless of storage temperature(P>0.05)compared with that at baseline(0 h,-0.56±1.51).④The BCR-ABL copy number of the overall sample only decreased significantly(P<0.05)at 48 h(2.93±1.59)and 72 h(2.79±1.42)compared with that at baseline(0 h,3.35±1.60)when stored at room temperature.The ABL copy number in the overall sample decreased significantly at 48 and 72 h(whether low and room temperature;P<0.05).However,no significant changes were noted in the BCR-ABL(P210)transcript levels after vibration for 3 h(-1.29±1.81),6 h(-1.24±1.72),and 12 h(-1.18±1.68;P>0.05)compared with that at baseline(0 h,-0.60±1.37).Conclusion Sample storage time,storage temperature,and vibration can interfere with the results of BCR-ABL and A

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 存储条件 运输 

分 类 号：R733.72[医药卫生—肿瘤]