STR-PCR定量检测供受者嵌合体方法的建立及临床应用

利用STRs的高度多态性等特性,用竞争性聚合酶链反应（PCR）结合聚丙烯凝胶电泳、染色和凝胶成像分析等技术,建立定量检测异基因造血干细胞移植后供受者嵌合体的方法,用无关个体的DNA混合实验进行可行性和准确性研究,并同时做染色体分带进行对照,以此基础上对2例非清髓异基因造血干细胞移植病人的植入情况进行动态检测。结果显示,扩增产物中供受者DNA含量的百分比同扩增前混合样本的比例呈显著直线相关,STR－PCR检测嵌合体方法简单、快速、可靠、敏感性高,所需标本量少且不受性别限制,是检测NAST供受者嵌合体的新方法。

QUANTITATIVE ASSESSMENT OF MIXED HEMATOPOIETIC CHIMERISM BY STR-PCR AFTER NONMYLOABLATIVE ALLOGENEIC STEM CELL TRANSPLANTATION

To establish a method for quantitative assessment of mixed hematopoietic chimerism by competitive polymerase chain reaction after nonmyeloablative allogeneic stem cell transplantation(NAST) using highly polymorphic short tandem repeats (STRs). The feasibility of this assay and the accuracy of quantitative results were tested using serial DNA mixtures of unrelated individuals. 2patients receiving NAST were analyzed by this method. Chromosomewas analyzed simultaneously. The results showed a linear correlation between the DNA added and the proportion found. Comparison of values obtained with chromosomeanalysis showed an excellent correlation with the STR PCR results. Taken together, this novel approach allows a rapid, sensitive, versatile, less blood , even with minuscule numbers of cells,and also sex unlimited.