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[用于血液系统恶性肿瘤患者嵌合体分析的多重短串联重复序列(STR)-PCR技术的开发及不同样本来源嵌合体的比较]

[Development of multiplex short tandem repeat (STR)-PCR for chimerism analysis in patients with hematological malignancies and comparison of chimerism in different sample sources].

作者信息

Taira Chiaki, Matsuda Kazuyuki, Takezawa Yuka, Ito Toshiro, Ishida Fumihiro, Hidaka Eiko, Kumagai Toshiko, Honda Takayuki

机构信息

Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto 390-8621, Japan.

出版信息

Rinsho Byori. 2011 Jan;59(1):24-30.

PMID:21404576
Abstract

Polymerase chain reaction analysis of short-tandem repeat (STR) markers (STR-PCR) has been used for chimerism testing to assess engraftment following hematopoietic stem cell transplantation (HSCT). We investigated the informativity of 7 STR loci (FGA, D5S818, SE33, TH01, VWF, PentaE, and D18S51) in 82 pre-HSCT DNA samples from 41 donor/recipient pairs and developed 2 multiplex STR-PCRs using VWF, SE33, and D18S51, D5S818 and FGA, respectively. The multiplex STR-PCRs could distinguish the recipients and donors in 92.7% of the cases. Dilution experiments using mixed DNA showed that the sensitivity of the multiplex STR-PCRs for detecting the minor population was 1-5%. To compare chimerism in different samples such as peripheral blood, mononuclear cells (MNC), and CD3-positive cells (CD3+), we investigated the relationship between the chimerisms at approximately day 30 post-HSCT and the interval from the day of HSCT to achievement of complete chimerism (CC) in 70 patients undergoing HSCT. CC was found in all samples of 54 patients at day 30 post-HSCT, and these samples showed CC thereafter. Eleven patients with mixed chimerism (MC) in all samples or in MNC and CD3+ showed CC at day 60-270 post-HSCT or persistent MC. The remaining 5 patients with MC in only CD3+ showed CC at day 30-60 post-HSCT. Taken together, MNC which can be separated easily may be a useful source for detecting patients who require longer time to achieve CC and those with high risk of graft failure.

摘要

短串联重复序列(STR)标记的聚合酶链反应分析(STR-PCR)已用于嵌合体检测,以评估造血干细胞移植(HSCT)后的植入情况。我们研究了41对供体/受体的82份HSCT前DNA样本中7个STR位点(FGA、D5S818、SE33、TH01、VWF、PentaE和D18S51)的信息性,并分别使用VWF、SE33以及D18S51、D5S818和FGA开发了2种多重STR-PCR。多重STR-PCR在92.7%的病例中能够区分受体和供体。使用混合DNA进行的稀释实验表明,多重STR-PCR检测少量细胞群体的灵敏度为1%-5%。为了比较不同样本(如外周血、单核细胞(MNC)和CD3阳性细胞(CD3+))中的嵌合情况,我们研究了70例接受HSCT患者在HSCT后约30天的嵌合情况与从HSCT日到实现完全嵌合(CC)的间隔时间之间的关系。54例患者在HSCT后30天所有样本中均出现CC,此后这些样本均显示为CC。11例在所有样本或MNC和CD3+中出现混合嵌合(MC)的患者在HSCT后60-270天出现CC或持续存在MC。其余仅在CD3+中出现MC的5例患者在HSCT后30-60天出现CC。综上所述,易于分离的MNC可能是检测需要更长时间实现CC的患者和移植失败高风险患者的有用来源。

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