二代测序技术监测急性淋巴白血病的克隆演变  

Next-generation genome sequencing technology in monitoring clonal evolution of relapsed acute lymphoblastic leukemia

作　　者：张英驰 章婧嫽 竺晓凡 

机构地区：[1]中国医学科学院血液病医院血液学研究所儿童诊疗中心,天津300020

出　　处：《中华实用诊断与治疗杂志》2016年第6期566-567,共2页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家重大科学研究计划项目(2012CB966603);天津市科技支撑计划项目(2ZCDZSY18100);国家自然科学基金青年科学基金(81400137)

摘　　要：目的采用二代测序技术监测复发儿童B细胞急性淋巴白血病(B cell acute lymphoblastic leukemia,B-ALL)的克隆演变。方法 20例B-ALL患儿的初诊和复发骨髓标本,分别提取全基因组DNA,通过多重PCR方法扩增免疫球蛋白重链基因(immunoglobulin heavy chain,IGH)互补决定区3(complementarity determining region 3,CDR3)序列并进行二代测序检测。结果 20例B-ALL患儿初诊骨髓标本IGH CDR3测序结果显示,17例有CDR3特异性序列,其中9例有1种CDR3特异性序列,7例有2种CDR3特异性序列,1例有3种CDR3特异性序列;复发骨髓标本IGH CDR3测序结果显示,16例有CDR3特异性序列,其中9例有1种CDR3特异性序列,7例有2种CDR3特异性序列;对初诊和复发标本CDR3序列进行比对分析,发现4例患儿出现克隆演变。结论采用二代测序技术可监测儿童B-ALL复发过程中的克隆演变。Objective To monitor the clonal evolution of pediatric relapsed B cell acute lymphoblastic leukemia（B-ALL）with next-generation sequencing（NGS）technology.Methods Twenty pediatric B-ALL samples were collected at initial diagnosis and relapse.Genomic DNA was extracted for PCR amplification of the immunoglobulin heavy chain（IGH）locus complementarity-determining region 3（CDR3）with subsequent NGS.Results The sequence and analysis results of IGH CDR3 of bone marrow samples from 20 initially diagnosed B-ALL patients showed leukemia-specific CDR3 sequences in 17,in which 9 patients were found one specific CDR3 sequences,7 were found two specific CDR3 sequences,and 1was found three specific CDR3 sequences.The sequence and analysis results of IGH CDR3 of relapsed bone marrow samples showed leukemia-specific CDR3 sequences in 16,in which 9 patients were found one specific CDR3 sequences,and 7 were found two specific CDR3 sequences.Through sequence alignment of specific CDR3 sequence between initially diagnosed and relapsed patients,clonal evolution was found in 4 patients.Conclusion NGS can be used for clonal evolution monitoring in pediatric B-ALL.

关 键 词：儿童白血病 二代测序技术 克隆演变 免疫球蛋白重链基因 

分 类 号：R733.71[医药卫生—肿瘤]