STR-PCR半定量检测嵌合体方法在异基因造血干细胞移植中的应用(1)
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[摘要] 目的:应用短串联重复序列-聚合酶链反应(STR-PCR)半定量方法检测异基因造血干细胞移植后供者细胞比例并探讨其临床意义。方法：给30例接受移植的患者行移植后嵌合体检测,采集外周血提取DNA,对9个STR位点行聚合酶链反应扩增,扩增完成后行聚丙烯酰胺凝胶电泳及硝酸银染色显影,再计算嵌合率。结果：30例患者均检测到供者细胞,24例达到完全嵌合(CC),6例形成混合造血(MC)。在移植后14 d,稳定植入的患者多数表现为CC。CC组的aGVHD发生率高于MC组,7例患者在复发前检测到嵌合率(donor cell,DC)下降,经相应免疫治疗后有3例达到CC。结论：监测异基因造血干细胞移植后嵌合率的动态变化过程,PCR-STR是一种简单经济的方法,可根据嵌合状态指导个体化免疫治疗,对患者的长期存活有重要意义。
[关键词] 短串联重复序列-聚合酶链反应； 异基因造血干细胞移植； 嵌合体分析
[中图分类号] R733.7; R730.54 [文献标识码] A [文章编号] 1671-7256(2011)02-0138-04
doi：10.3969/j.issn.1671-7256.2011.02.005
Semi-quantitative analysis of chimerism after allogeneic
stem cell transplantation using STR-PCR
LIU Jing, DING Jia-hua
(Department of Hematology, Zhongda Hospital, Southeast University， Nanjing 210009, China)
[Abstract] Objective: To investigate the value of semi-quantitative analysis of the ratio of donor cells after allogeneic hematopoietic stem cell transplantation(allo-HSCT) using short tandem repeat-polymerase chain reaction (STR-PCR). Methods： Among 30 patients receiving allo-HSCT were evaluated, DNA was extracted from peripheral blood and nine STR loci were amplified followed by polyacrylamide gels, silver staining,and then the engomphosis rates were calculated. Results: Donor cells could be detected from all of 30 cases, 24 cases formed complete chimerism(CC) and 6 cases formed mixed chimerism(MC). Most patients with stable engraftments showed CC on day+14. The incidence of aGVHD in group CC was significantly higher than that in group MC. Before the time of relapse, STR-PCR indicated a decrease of donor cell(DC) in 7 patients, and 3 cases were of complete chimerism by corresponding immunotherapy. Conclusions： STR-PCR provide a simple and economic approach for monitoring the dynamic process of donor chimerism. Individual immunotherapy based on chimerism is important for patient long time survial.
[Key words] short tandem repeat-polymerase chain reaction; allogeneic hematopoietic stem cell transplantation; analysis of chimerism
异基因造血干细胞移植现今已经成为临床上各种恶性及非恶性血液病治疗的一项重要措施,供者细胞植入受者体内后供者细胞及受者细胞含量情况是动态变化的,需要进行动态的检测。检测结果有助于判断移植是否成功,及预测移植物抗宿主病的发生与疾病的复发,并指导下一步免疫干预治疗的应用[1]。短串联重复序列(STR)是由2～6个碱基对组成的一种遗传标记,已在移植后嵌合体检测中广泛使用。传统的STR-PCR检测方法敏感性多大于5%,但当供受者性别相同时,无法计算供者细胞的嵌合率 ......
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