Suitable Molecular Genetic Methods for the Monitoring of Cell Chimerism

H. Cechova, Lucie Pavlátová, Monika Leontovycova, M. Vraná
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Abstract

The molecular analysis of individual hematopoietic chimerism at a defined time after allogeneic hematopoietic stem cell transplantation represents an important non-specific marker of posttransplant course. The monitoring of its dynamic allows the identification of patients at a high risk of relapse. A variety of methods are used for the monitoring of cell chimerism. It is necessary to use sensitive molecular genetic methods for early detection of the autologous hematopoiesis. Quantitative multiplex real-time polymerase chain reaction (PCR) analysis can serve as a very sensitive (0.01 – 0.1%), relatively quick, and inexpensive method to detect < 1% of minor genotype. With an increasing ratio of minor genotype ( > 1%), it is more suitable to use short tandem repeats (STRs) for its analysis. Based on the differences in recipient/donor pair genotypes, at least two suitable informative polymorphisms located at different chromosomes can be selected. The combination of methods is appropriate, and the choice of the used method depends on the patient ’ s actual chimerism status. The cohort of 207 patients monitored at the Institute of Hematology and Blood Transfusion was divided into three subgroups according to their chimerism status (complete chimerism (CC), microchimerism, mixed chimerism (MC)) 3 years after allogeneic hematopoietic stem cell transplantation (allo-HSCT). A significant difference in the 3-year survival and 3-year relapse rates in all three subgroups was found.
适用于细胞嵌合体监测的分子遗传学方法
同种异体造血干细胞移植后一定时间内个体造血嵌合的分子分析是移植后病程的重要非特异性标志物。监测其动态可以识别复发风险高的患者。用于监测细胞嵌合的方法多种多样。采用灵敏的分子遗传学方法对自体造血进行早期检测是必要的。定量多重实时聚合酶链反应(PCR)是一种非常灵敏(0.01 - 0.1%)、相对快速、廉价的检测< 1%小基因型的方法。随着小基因型比例的增加(> %),更适合使用短串联重复序列(STRs)进行分析。根据受体/供体对基因型的差异,可以选择至少两个位于不同染色体上的合适信息多态性。方法的组合是适当的,所使用的方法的选择取决于患者的实际嵌合状态。在血液学和输血研究所监测的207例患者队列根据其嵌合状态(完全嵌合(CC),微嵌合,混合嵌合(MC))在同种异体造血干细胞移植(alloo - hsct)后3年分为三个亚组。三个亚组的3年生存率和3年复发率均有显著差异。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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