短串联重复序列监测异基因造血干细胞移植后的植入状态及其演变过程

Implantation status and its variation process of allogeneic peripheral blood stem cell transplantation monitored by short tandem repeat sequence

目的:监测和观察异基因造血干细胞移植后的植入状态及其演变过程。方法:于2003-01/2004-12在中国造血干细胞库深圳分库采集完成人类白细胞抗原配型的6对供受者标本。采用荧光标记复合扩增短串联重复序列检测技术,对6对供受者标本移植前后的系列血样进行D8S1179,D21S11,D18S51,D5S818,D13S317,D7S820,D3S1358,vWA,FGA共9个短串联重复序列位点和Amelogenin性别位点的检测,找出供受者间的差异基因,通过移植后不同时间段患者基因型的转变情况,判断供者细胞是否植入以及嵌合体类型。结果:供受者6对样本均获得满意的9个短串联重复序列位点和1个性别位点的分型结果。6对移植标本短串联重复序列差异基因由受者型向供者型转化的时间为14～23d。6对样本间存在明显差异。第3对、第5对在第14天已为供者完全嵌合状态,而第1对在第16天、第4对在第18天尚为供受者混合嵌合体。但6对样本都是一个从混合嵌合向完全嵌合发展的良好趋势。结论:①6对移植标本短串联重复序列差异基因由受者型向供者型转化的时间为14～23d,6对样本都有从混合嵌合向完全嵌合发展的良好趋势。②采用荧光标记复合扩增短串联重复序列方法可精确地测量聚合酶链反应产物的数量,描述造血干细胞的植入程度及植入的整个演变过程。

AIM： To monitor and observe the implantation status and its variation process of allogeneic peripheral blood stem cell transplantation （allo-PBSCT）. METHODS： Six pairs of recipient-donors specimen matched with human leucocyte antigen （HLA） were collected from Shenzhen branch of China hemopoietic stem cell bank from January 2003 to December 2004. Series of blood sample of the six pairs of recipient-donor specimen before and after transplantation were detected on nine STR sequence locus, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, D3S1358, vWA, FGA and Amelogenin sex site by fluorescence labeling compound-amplification short tandem repeat （STR） detection technique to find the different gene among recipient-donors. To judge whether the cells of donors was transplanted and the type of chimera through the change condition of the gene in patients in different time period after transplantation. RESULTS： All the six pairs of recipient-donors specimens got the satisfactory typing result of 9 STR sequence site and 1 sex site. The conversion time of different gene of the six pairs of transplantation specimens STR sequence from recipient to donor was 14-23 days. There were significant differences among the six pairs of specimens. The third pair and fifth pair at 14^th day had been complete gomphosis donor status already. The first pair at 16^th day and the fourth pair at 18^th day had been still recipient-donor mixture chimera. However, the six pairs of exponent all had a well development trend from mixture chimera to complete gomphosis. CONCLUSION： ①The conversion time of different gene of the six pairs of transplantation specimens STR sequence from recipient to donor is 14- 23 days, and the six pairs of exponent all have a well development trend from mixture chimera to complete gomphosis. ② Using the fluorescence labeling compound-amplification STR sequence technique, the quantity of polymerase chain reaction （PCR） product can be measured precisely, and the implanted level and the whole implanted variation process of hemopoietic stem cells can be described.