BCR-ABL1候选参考物质的研制及测量不确定度评定  

Development of BCR-ABL1 candidate reference material and evaluation of measurement uncertainty

作　　者：于书平 袁丹丹 崔明[1] 景蓉蓉[1] 王惠民[1] YU Shuping;YUAN Dandan;CUI Ming;JING Rongrong;WANG Huimin(Department of Clinical Laboratory,the Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南通大学附属医院医学检验科

出　　处：《检验医学》2019年第8期752-757,共6页Laboratory Medicine

基　　金：江苏省青年医学重点人才项目(QNRC2016687)

摘　　要：目的构建慢性粒细胞白血病(CML)融合基因断裂点簇集区-埃布尔森小鼠白血病病毒癌基因1(BCR-ABL1)候选参考物质。方法采用分子克隆技术构建含有BCR-ABL1基因的重组质粒,测序验证后采用紫外分光光度法初步测定浓度,并采用数字聚合酶链反应(dPCR)进行浓度定值,实时荧光定量聚合酶链反应(RQ-PCR)进行均匀性和稳定性研究,最终应用《测量不确定度表示指南》评定测量不确定度。结果质粒候选参考物质拷贝数浓度为(2.50±0.35)×10^6拷贝/μL,有较好的均匀性和稳定性。结论建立了制备BCR-ABL1候选参考物质的方法,构建了均匀稳定的BCR-ABL1候选参考物质。Objective To develop the breakpoint cluster region-abelson murine leukemia viral oncogene homolg 1(BCR-ABL1)fusion gene candidate reference material for chronic myeloid leukemia(CML).Methods A plasmid containing the BCR-ABL1 fusion gene was constructed by molecular cloning technology,and its concentration was quantified by ultraviolet spectrophotometry and digital polymerase chain reaction(PCR)following sequence validation.The homogeneity and stability were evaluated by real-time quantitative PCR(RQPCR).The uncertainty was evaluated according to the Guide to the Expression of Uncertainty in Measurement.Results The copy concentration of plasmid candidate reference material was(2.50±0.35)×10^6 copies/μL.The homogeneity and stability of plasmid were good.Conclusions The method for constructing BCR-ABL1 candidate reference material and a homogeneous and stable BCR-ABL1 candidate reference material have been established.

关 键 词：断裂点簇集区-埃布尔森小鼠白血病病毒癌基因1 参考物质 测量不确定度 数字聚合酶链反应 紫外分光光度法 

分 类 号：R446.1[医药卫生—诊断学]