Detection of minimal residual disease B cell lymphoma by a PCR-mediated RNase protection assay.
Detection of minimal residual disease B cell lymphoma by a PCR-mediated RNase protection assay.
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通过 PCR 介导的 RNase 保护测定检测微小残留病 B 细胞淋巴瘤。
作者：
T. Kurokawa;T. Kinoshita;T. Ito;H. Saito;T. Hotta
为了检测和监测B细胞淋巴瘤患者骨髓（BM）或外周血（PB）中的肿瘤细胞，我们使用PCR介导的RNase保护试验来鉴定互补决定区（CDR）-III基因重排。该方法既不需要确定核苷酸序列，也不需要构建肿瘤特异性寡核苷酸探针或引物。使用B细胞系以及临床样品的该测定的灵敏度显示在10（4）-10（5）个正常细胞的背景中有一个肿瘤细胞。使用这种方法，我们检查了31例B细胞淋巴瘤患者，其中初始肿瘤组织的IgH重排被Southern印迹分析证实。31例患者中的20例（65%）能够使用该测定进行分析。在所有可评价样本中，通过本试验确认了常规形态学检查或表面标志物分析中明显存在的BM和PB中的肿瘤细胞。此外，我们检测了5例患者的BM或PB中的肿瘤细胞，其中常规方法未发现肿瘤细胞。PCR介导的RNase保护试验具有灵敏、快速、简便等优点，可用于B细胞淋巴瘤微小残留病（MRD）的检测。
To detect and monitor tumor cells in the bone marrow (BM) or peripheral blood (PB) of patients with B cell lymphoma, we used the PCR-mediated RNase protection assay to identify the complementarity determining regions (CDR)-III gene rearrangement. This method required neither determination of nucleotide sequences nor construction of tumor-specific oligonucleotide probes or primers. The sensitivity of this assay using B cell lines as well as clinical samples revealed one tumor cell in a background of 10(4)-10(5) normal cells. Using this assay we examined 31 patients with B cell lymphoma in whom an IgH rearrangement of initial tumor tissues was confirmed by Southern blot analysis. Twenty of 31 (65%) patients were able to be analyzed using this assay. Tumor cells in the BM and PB evident on routine morphological examination or surface marker analysis were confirmed by this assay in all the evaluable samples. Moreover, we detected tumor cells in the BM or PB of five patients in whom no tumor cells were identified by conventional methods. The PCR-mediated RNase protection assay is useful to detect minimal residual disease (MRD) in B cell lymphoma because it is highly sensitive, rapid and simple.