Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction
Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction
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DOI：
10.1038/sj.bmt.1704764
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发表时间：
2005-02-01
影响因子：
4.8
通讯作者：
Ellard, S
中科院分区：
文献类型：
--
作者：
Harries, LW;Wickham, CL;Ellard, S
异基因骨髓移植（BMT）与骨髓烧蚀预处理是治疗造血系统恶性肿瘤的选择。非清髓性干细胞移植的使用使得以前由于年龄或其他疾病而不适合BMT的患者得到治疗。这些减少的调节机制允许血液和骨髓中的一些受体细胞最初持续存在（造血嵌合现象）。需要监测供体和受体细胞的相对比例，以评估手术的成功，预测随后的排斥或即将复发，并指导供体淋巴细胞输注的使用。我们提出了一种定量实时PCR方法，用于使用TaqMan（TM）测量造血嵌合体。这种方法利用单核苷酸多态性（SNP）的存在来区分患者或供体来源的细胞。我们设计并验证了一组7个等位基因特异性探针，以量化患者和供体细胞在12个患者和供体对的造血群体中的贡献。我们已经比较了这种方法的性能与现有的方法，并证明它是优越的准确性和灵敏度的上级。使用更敏感和准确的技术允许早期干预，以改善临床结果。
Allogeneic bone marrow transplantation (BMT) with marrow ablative conditioning is the treatment of choice for haematopoietic malignancies. The use of nonmyeloablative stem cell transplants has allowed the treatment of patients previously ineligible for BMT because of age or other disease. These reduced conditioning regimes allow the persistence initially of some recipient cells in the blood and bone marrow ( haematopoietic chimaerism). Monitoring of the relative proportion of donor and recipient cells is required to assess the success of the procedure, to predict subsequent rejection or impending relapse and to guide the use of donor lymphocyte infusions. We present a quantitative real-time PCR approach for the measurement of haematopoietic chimaerism using the TaqMan(TM). This approach exploits the presence of single-nucleotide polymorphisms ( SNPs) to distinguish cells of patient or donor origin. We have designed and validated a panel of seven allele-specific probes to quantify the contribution of patient and donor cells in the haematopoietic population from 12 patient and donor pairs. We have compared the performance of this approach with an existing method and proved it to be superior in both accuracy and sensitivity. The use of more sensitive and accurate techniques permits earlier intervention for improved clinical outcome.