适合移植后嵌合物定量分析的信息STR位点谱的建立及其应用  

Short Tandem Repeat Panel Settled for Quantitative Chimerism Analysis Following Allogeneic Stem Cell Transplantation and Its Application

作　　者：杨颖[1] 吴文[2] 王玲玲[1] 陈和平[1] 朱自严[1] 

机构地区：[1]上海市血液中心、上海输血研究所,上海200051 [2]上海市瑞金医院血液科,上海200025

出　　处：《中国实验血液学杂志》2009年第3期706-712,共7页Journal of Experimental Hematology

基　　金：上海市科委资助浦江人才计划(编号06PJ14080);上海卫生局资助项目(编号2008056)

摘　　要：本研究选取8个STR位点作为嵌合物定量分析的信息STR位点谱,评估其作为异基因造血干细胞移植(HSCT)后移植物嵌合状态的定量监控指标的作用。采集15对移植供受者血样,提取DNA,分别合成标记D3S3045、D4S2366、D4S2639、D5S818、D13S317、D18S1002、D20S481、D22S689的引物,建立PCR扩增体系,在ABI3100仪上检测这些位点的扩增片段,筛选得到理想的信息位点;制备梯度供者/受者DNA混合品和移植后不同日期采集的标本,进行信息位点的检测;获取峰高或峰面积进行移植嵌合率的计算。结果显示,根据梯度混合DNA的结果获得标准曲线,计算得到的嵌合率和配制浓度中的嵌合比例基本一致;用峰高和峰面积同时计算,两者基本一致;成功获得了15例移植后病人的嵌合状态变化数据,并帮助诊断了1例复发病人。结论:本研究建立的信息STR位点谱及检测方法,可以较准确地定量监控嵌合状态,成功用于临床干细胞移植工作,与商品化试剂相比,更便宜,在使用上更灵活。To evaluate the roles of 8 short tandem repeats（STR） loci as STR panel in quantitative analysis of chimerism following transplantation, the primers were synthesized and marked with different dyes for D3S3045, D4S2366, D4S2639,DSSS18,D13S317, D18S1002, D20S481 and D22S689. The blood samples of 15 cases received allogeneic stem cell transplantation were collected before and after transplantation, then DNA was extracted and amplified with these primers, and was further analysed under ABI Genetic Analyser 3100 to select suitable informative STR locus. Donor/recipient dilution series were prepared to get standard curves in selected loci, the DNAs extracted at different days after transplantation were used to quantitatively analyze the chimerism in patients according to the values of peak area or peak height of fluorensent signals. The standard curves can be used to calculate the chimerism by plottting the respective R/D quotient value against the percentage of recipient DNA. The results indicated that the calculated chimerism was in concordance with the donor/recipient dilution. The STR panel succeeded in identifying at least one informative marker and quantitative monitoring the chimerism after HSCT in 15 donor-recipient pairs and a relapsed case was diagnosed. It is concluded that the STR panel and its detection method can accurately and quantitatively monitor the chimerism after allogeneic HSCT, which is more economical and flexible than using commercial kits.

关 键 词：STR位点谱 嵌合状态 定量分析 异基因造血干细胞移植 

分 类 号：R730.5[医药卫生—肿瘤] R457.7[医药卫生—临床医学]