摘要
目的 建立一种利用多核苷酸多态性高通量测序(MNPseq)分析异基因造血干细胞移植后嵌合状态的新方法,并探讨其可行性及优越性.方法 筛选100个MNP片段,采用高通量测序技术,通过模拟嵌合样本和临床移植后样本,与STR法、融合基因定量检测和流式细胞术微小残留病检测进行对比,验证方法的准确性和敏感性.结果 MNPseq的准确性和敏感性均优于STR法,其中敏感性为0.01%,较STR法敏感约100倍;MNPseq可以进一步区分STR完全嵌合的42份样本,且经Cutoff值校正后,与融合基因定量检测结果相关;MNPseq可以纠正因为影子峰所造成的STR法的假阳性,并且可以用于检测缺乏供者和(或)患者移植前信息的嵌合体标本.结论 基于高通量测序的MNPseq分析是一种更加准确和敏感的嵌合体检测方法,而且解决了缺乏移植前信息无法检测嵌合体的问题,具有极高的临床应用价值.
Objective To establish a new method for chimerism analysis after allogeneic hematopoietic stem cell transplantation by using multiple nucleotide polymorphism sequencing(MNPseq),and to explore its feasibility and superiority.Methods One hundred MNP fragments were screened and chimeric analysis was performed by high-throughput sequencing technology.The accuracy and sensitivity of the method were verified by simulating chimeric samples and post-transplant samples and comparing them with short tandem repeat(STR)analysis,fusion gene quantitative detection and flow cytometry for minimal residual disease.Results The accuracy and sensitivity of MNPseq were better than those of STR,in which the sensitivity could reach 0.01%,about 100 times more sensitive than STR.MNPseq could further distinguish 42 STR fully chimeric samples,and after corrected by cutoff value,it was correlated with the quantitative detection of fusion gene.MNPseq could correct false positive of STR caused by the shadow peak,and could be used to detect chimeric samples lacking pre-transplant information from donors and recipients.Conclusion MNPseq analysis based on high-throughput sequencing is a more accurate and sensitive chimerism detection method,and it solves the problem that chimerism cannot be detected due to the lack of pre-transplant information,which has extremely high clinical application value.
作者
吕晓东
邹喆
彭海
范瑞华
宋永平
Lyu Xiaodong;Zou Zhe;Peng Hai;Fan Ruihua;Song Yongping(Central Lab,the Affiliated Cancer Hospital of Zhengzhou University,Henan Cancer Hospital,Zhengzhou450008,China;Institute of Systematic Biology,Jianghan University,Wuhan430056,China;Departmentof Hematology,the Affiliated Cancer Hospital of Zhengzhou University,Henan Cancer Hospital,Zhengzhou450008,China)
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2019年第8期662-666,共5页
Chinese Journal of Hematology
基金
河南省自然科学基金(182300410371)
河南省科技攻关项目(192102310051).
关键词
多核苷酸多态性
高通量测序
嵌合体
异基因造血干细胞移植
Multiple nucleotide polymorphism
high-throughput sequencing
Chimerism
Allogeneic hematopoietic stem cell transplantation
