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Quantitative assessment of hematopoietic chimerism by quantitative real-time polymerase chain reaction of sequence polymorphism systems after hematopoietic stem cell transplantation
Qin Xiao-ying; Li Guo-xuan; Qin Ya-zhen; Wang Yu; Wang Feng-rong; Liu Dai-hong; Xu Lan-ping; Chen Huan; Han Wei; Wang Jing-zhi; Zhang Xiao-hui; Li Jin-lan; Li Ling-di; Liu Kai-yan; Huang Xiao-jun
关键词sequence polymorphism single nucleotide polymorphism real-time PCR hematopoietic stem cell transplantation chimerism
刊名CHINESE MEDICAL JOURNAL
2011-08-05
DOI10.3760/cma.j.issn.0366-6999.2011.15.011
124期:15页:2301-2308
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Medicine, General & Internal
研究领域[WOS]General & Internal Medicine
关键词[WOS]BONE-MARROW-TRANSPLANTATION ; SINGLE-NUCLEOTIDE POLYMORPHISMS ; CLINICAL-BIOLOGICAL SITUATIONS ; MULTIPLEX PCR AMPLIFICATION ; MINIMAL RESIDUAL DISEASE ; TANDEM REPEAT-MARKERS ; MIXED CHIMERISM ; CAPILLARY-ELECTROPHORESIS ; ADOPTIVE IMMUNOTHERAPY ; MICROSATELLITE MARKERS
英文摘要

Background Analysis of changes in recipient and donor hematopoietic cell origin is extremely useful to monitor the effect of hematopoietic stem cell transplantation (HSCT) and sequential adoptive immunotherapy by donor lymphocyte infusions. We developed a sensitive, reliable and rapid real-time PCR method based on sequence polymorphism systems to quantitatively assess the hematopoietic chimerism after HSCT.

Methods A panel of 29 selected sequence polymorphism (SP) markers was screened by real-time PCR in 101 HSCT patients with leukemia and other hematological diseases. The chimerism kinetics of bone marrow samples of 8 HSCT patients in remission and relapse situations were followed longitudinally.

Results Recipient genotype discrimination was possible in 97.0% (98 of 101) with a mean number of 2.5 (1-7) informative markers per recipient/donor pair. Using serial dilutions of plasmids containing specific SP markers, the linear correlation (r) of 0.99, the slope between -3.2 and -3.7 and the sensitivity of 0.1% were proved reproducible. By this method, it was possible to very accurately detect autologous signals in the range from 0.1% to 30%. The accuracy of the method in the very important range of autologous signals below 5% was extraordinarily high (standard deviation <1.85%), which might significantly improve detection accuracy of changes in autologous signals early in the post-transplantation course of follow-up. The main advantage of the real-time PCR method over short tandem repeat PCR chimerism assays is the absence of PCR competition and plateau biases, with demonstrated greater sensitivity and linearity. Finally, we prospectively analyzed bone marrow samples of 8 patients who received allografts and presented the chimerism kinetics of remission and relapse situations that illustrated the sensitivity level and the promising clinical application of this method.

Conclusion This SP-based real-time PCR assay provides a rapid, sensitive, and accurate quantitative assessment of mixed chimerism that can be useful in predicting graft rejection and early relapse. Chin Med J 2011;124(15):2301-2308

语种英语
WOS记录号WOS:000294584100011
项目编号30725038
资助机构National Outstanding Young Scientist&prime ; s Foundation of China
引用统计
被引频次:9[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/61096
专题北京大学第二临床医学院_血液科
医学人文研究院/公共教学部_哲学与社会科学系
作者单位Peking Univ, Peoples Hosp, Inst Hematol, Beijing Key Lab Hematopoiet Stem Cell Transplanta, Beijing 100044, Peoples R China
推荐引用方式
GB/T 7714
Qin Xiao-ying,Li Guo-xuan,Qin Ya-zhen,et al. Quantitative assessment of hematopoietic chimerism by quantitative real-time polymerase chain reaction of sequence polymorphism systems after hematopoietic stem cell transplantation[J]. CHINESE MEDICAL JOURNAL,2011,124(15):2301-2308.
APA Qin Xiao-ying.,Li Guo-xuan.,Qin Ya-zhen.,Wang Yu.,Wang Feng-rong.,...&Huang Xiao-jun.(2011).Quantitative assessment of hematopoietic chimerism by quantitative real-time polymerase chain reaction of sequence polymorphism systems after hematopoietic stem cell transplantation.CHINESE MEDICAL JOURNAL,124(15),2301-2308.
MLA Qin Xiao-ying,et al."Quantitative assessment of hematopoietic chimerism by quantitative real-time polymerase chain reaction of sequence polymorphism systems after hematopoietic stem cell transplantation".CHINESE MEDICAL JOURNAL 124.15(2011):2301-2308.
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