B细胞非霍奇金淋巴瘤IgH基因重排检测及凝胶扫描技术应用的初步研究  

Detection of IgH gene rearrangement in B cell non Hodgkin′s lymphoma and application of gel scan method

作　　者：张晓飞[1] 周韧[1] 毛峥嵘[1] 骆利康[2] 胡孟钧 杨水友[1] 李斐铭 

机构地区：[1]浙江大学病理与法医学研究所和浙江大学医学院环境基因组研究中心 [2]浙江省衢州市人民医院病理科 [3]诸暨市人民医院病理科 [4]东阳市人民医院

出　　处：《中华病理学杂志》2004年第6期513-517,共5页Chinese Journal of Pathology

基　　金：浙江省科技厅科研基金 (0 0 1110 42 5 );浙江省卫生厅科研基金 (2 0 0 2ZX0 10 )资助

摘　　要：目的　采用凝胶扫描技术 ,力求对采用聚合酶链反应 (PCR)检测非霍奇金淋巴瘤(NHL)抗原受体基因重排结果分析时建立量化标准 ,以利于临床医师应用时客观判断 ,并为其筛选随访病例提供依据。方法　用PCR对 96例B NHL分别采用IgHFR3A及FR2A检测IgH基因克隆性重排。 6 5例经IgHFR3APCR已证实为IgH基因克隆性重排B NHL及 8例良性病变淋巴组织和 5例正常人外周血单核细胞 ,IgHFR3APCR产物 8%变性聚丙烯酰胺凝胶电泳银染后图像行凝胶扫描 ,绘制曲线并计算h1/h2比值。结果　 (1)采用FR3A引物 ,克隆性IgH基因重排在 96例B NHL中检测率为 6 8% ,采用FR2A引物 ,检测率为 6 1% ,结合FR3A、FR2A引物 ,总检测率为 83%。 (2 )凝胶扫描曲线示 6 5例B NHL的h1/h2均〉3,而 5例正常人外周血单核细胞曲线均表现为钟型 ,8例良性病变淋巴组织h1/h2比值均 <1 5。结论　非霍奇金淋巴瘤抗原受体基因重排检测的凝胶扫描分析曲线中 ,h1/h2峰高比值至少 >3可提示为IgH基因克隆性重排 ,<1 5可能提示IgH基因多克隆重排。比值介于 1 5和 3之间 ,提示有随访意义。联合FR3A ,FR2A引物 ,可提高B NHLIgH基因克隆性重排的检测率。Objective To develop a protocol for gene rearrangement study in non Hodgkin′s lymphoma (NHL) by PCR directed gel scan method and to set up quantitative criteria for IgH gene rearrangement which can be applied in the follow up of lymphoma patients. Methods IgH gene rearrangement studies were carried out in 96 cases of B cell NHL. The detection rate of clonality was evaluated. Sixty five cases of IgH gene rearranged cases proven by FR3A directed PCR and PAGE and 8 cases of benign lymphoid tissues (5 cases of reactive lymphoid hyperplasia, 3 cases of chronic tonsillitis), 5 cases of normal peripheral blood mononuclear cells were analyzed by gel scan method and the proportion of h1/h2 (heights of peak1 and peak2 of gel scan)was calculated. Results The detection rate of IgH gene clonality was up to 68% using primer FR3A in the 96 B cell NHL cases. The detection rate was up to 61% using primer FR2A. With a combination of primers FR3A and FR2A, the detection rate increased to 83%. Gel scan curve showed that the value of h1/h2 was greater than 3 in all the 65 cases with IgH gene rearranged. In the 8 benign lymphoid tissue cases showed h1/h2<1 5, 5 cases with normal peripheral blood mononuclear cells showed a bell shaped curve.Conclusions In the gel scan curve of gene rearrangement studies in non Hodgkin′s lymphoma samples, the value of h1/h2 greater than 3 represents a true clonal proliferation. The peaks with relative heights less than 1 5 may not be significant and likely represent polyclonal cell population. A value between 1 5 and 3 however requires clinical follow up. The success rate of rearrangement studies in B cell NHL can be increased by using a combination of primers FR3A and FR2A.

关 键 词：NHL IGH基因重排 克隆性 非霍奇金淋巴瘤 比值 FR 良性病变 曲线 结论 提示 

分 类 号：R733.1[医药卫生—肿瘤]