Thiede C, Bornhäuser M, Oelschlägel U, Brendel C, Leo R, Daxberger H, Mohr B, Florek M, Kroschinsky F, Geissler G, Naumann R, Ritter M, Prange-Krex G, Lion T, Neubauer A, Ehninger G
Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus der Technischen Universität, Dresden, Germany.
Leukemia. 2001 Feb;15(2):293-302. doi: 10.1038/sj.leu.2401953.
Sequential analysis of chimerism after allogeneic blood stem cell transplantation (BSCT) has been shown to be predictive for graft failure and relapse. We have explored the impact of a novel approach for the quantitative determination of chimerism using a commercial PCR assay with multiplex amplification of nine STR-loci and fluorescence detection. The feasibility was studied in 121 patients transplanted from related or unrelated donors. Follow-up investigation was performed in 88 patients. Twenty-eight of these patients had received a transplantation after dose-reduced conditioning therapy. Results were compared to data obtained by FISH analysis in a subgroup of patients receiving grafts from sex-mismatched donors. The analysis was possible in all patients, the median number of informative alleles was 4 (range 1-8) compared to 7 (range 1-9) in the related and unrelated situation, respectively. A good correlation was seen in 84 samples from 14 patients analyzed in parallel with STR-PCR and FISH. Decreasing values of donor chimerism were detected prior to or concomitantly with the occurrence of graft failure and relapse of disease in all patients investigated prospectively. Using FACS-sorted material, eg peripheral blood CD34+ cells, the assay permitted the detection of residual recipient cells with high sensitivity (down to one CD34+ Kasumi cell in 40,000 normal WBC). Evaluation of the inter-laboratory reproducibility revealed that in 20 samples analyzed in three different centers, the median coefficient of variation was 2.1% (range 0.7-9.6%). Taken together, the results support the use of the test as a valuable tool in the follow-up of patients undergoing allogeneic BSCT. In cases lacking PCR-detectable disease-specific gene products, this assay may represent an alternative to recently established real-time PCR methods.
异体造血干细胞移植（BSCT）后嵌合体的序贯分析已被证明可预测移植失败和复发。我们探索了一种新方法的影响，该方法使用一种商业PCR检测法对9个STR位点进行多重扩增并进行荧光检测，以定量测定嵌合体。在121例接受相关或无关供体移植的患者中研究了其可行性。对88例患者进行了随访调查。其中28例患者在接受了剂量降低的预处理后进行了移植。将结果与在接受性别不匹配供体移植的患者亚组中通过FISH分析获得的数据进行比较。所有患者均能进行分析，信息性等位基因的中位数在相关和无关情况下分别为4（范围1 - 8）和7（范围1 - 9）。在14例患者的84个样本中，同时进行STR - PCR和FISH分析，结果显示出良好的相关性。在所有前瞻性研究的患者中，在移植失败和疾病复发之前或同时检测到供体嵌合体值下降。使用FACS分选的材料，如外周血CD34 +细胞，该检测法能够高灵敏度地检测残留的受体细胞（低至40,000个正常白细胞中有一个CD34 + Kasumi细胞）。实验室间重复性评估显示，在三个不同中心分析的20个样本中，变异系数中位数为2.1%（范围0.7 - 9.6%）。综上所述，这些结果支持将该检测法用作异体BSCT患者随访中的一种有价值工具。在缺乏PCR可检测的疾病特异性基因产物的情况下，该检测法可能是最近建立的实时PCR方法的一种替代方法。
