摘要
目的 为了准确地识别骨髓移植物的植入状态 ,探讨PCR扩增短串联重复序列 (shorttandemrepeat,PCR STR)在亲缘或无关供者骨髓移植的预后及白血病复发中的预警作用。方法 建立荧光标记PCR STR等位基因分析技术 ,采用单克隆磁珠提取DNA ,四色荧光标记 ,于移植前、移植后 7天～ 6个月不等时间 ,采集供、受者血液 (2例回顾性研究患者刮取口腔粘膜脱落细胞作为术前样本 )DNA作PCR STR分析。结果 ① 12例患者骨髓移植后 10例为完全供者型 ,其中同胞捐髓 8例 ,HLA全相合 7例 ,半相合 1例 ;无关供者捐髓 1例 ,HLA全相合 ,母亲供髓 1例 ,HLA Ⅰ全相合 ,HLA Ⅱ半相合。 10例受者术后出现Ⅰ度移植物抗宿主病 (GVHD) ,STR均完全和持续表达供者基因型 ,追踪观察 3～ 2 5个月 ,预后良好。② 12例中 1例由嵌合型转变为完全供者型 ,系同胞捐髓 ,HLA Ⅰ半相合 ,HLA Ⅱ全相合 ;术后第 30天PCR STR表达供、受者双方基因型 ,第 6 0天完全转变为供者基因型 ,而自身的基因在外周血中消失 ,预后良好。③ 12例中 1例为持续未植入 ,HLA半相合同胞捐髓 ,术后虽然有血液学和临床情况的改善 ,但移植后 7,14,2 1天STR分析始终仅显示受者基因型 ,移植后 4个月死亡。结论 基于分子水平的多位点PCR
Objective In order to determine the status of engraftment following allogeneic bone marrow transplantation (allo BMT) and predict the outcome, short tandem repeats (STR) were analysed by fluorescence labeling polymerase chain reaction (PCR). Method Fluorescence labeling PCR detecting STR 9 loci was established. DNA was extracted by monoclonal magnetic beads from recipients peripheral blood nucleated cells and labeled with four color fluorescences before and from 7 days to six months after allogeneic bone marrow transplantation.Results ①STR and clinical feature of 12 patients after allogeneic bone marrow transplantation were analysed. In the ten patients with full donor chimeras, 8 received marrow grafts from HLA identical sibling donors, 1 from HLA identical unrelated donor and 1 from HLA haploidentical mother. All of the 10 patients had durable engraftment and survived disease freely for 3～25 months of follow up.②One patient received a HLA AB loci mismatched graft and his STR showed mixed chimeras at day 30, and converted into full donor chimeras at day 60 after transplantation. ③One patient received HLA haploidentical sibling donor stem cells failed to be engrafted and died 4 months later. Conclusion Analysis of multi loci of STR after allo BMT provided a real indication of engraftment. Mixed chimeras after transplantation suggested that donor lymphocytes or bone marrow cells be re infused to enhance the conversion from mixed chimeras to full donor chimeras. For the documentation of engraftment,analysis of STR was better than any other available methods.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2001年第8期418-422,共5页
Chinese Journal of Hematology
