IgH基因单克隆重排检测及其在B细胞性非霍奇金淋巴瘤诊断中的应用  

Detection of monoclonal immunoglobulin heavy chain gene rearrangements and its application in diagnosis of B-cell non-Hodgkin lymphoma

作　　者：李银珍[1] 王芳[1] 邵琼[1] 张旭[1] 邓玲[1] 汤涛[1] 张晓[1] 吴秋良[2] 

机构地区：[1]华南肿瘤学国家重点实验室中山大学肿瘤防治中心分子诊断科,广东广州510060 [2]华南肿瘤学国家重点实验室中山大学肿瘤防治中心病理科,广东广州510060

出　　处：《中国病理生理杂志》2012年第11期1994-1998,共5页Chinese Journal of Pathophysiology

基　　金：中山大学肿瘤防治中心单病种科研基金(No.303040736005)

摘　　要：目的:建立准确可靠、操作性强、适用于临床实际工作的免疫球蛋白重链(immunoglobulin heavychain,IgH)基因单克隆重排检测方法,用于B细胞性非霍奇金淋巴瘤(B-cell non-Hodgkin lymphoma,B-NHL)的辅助诊断。方法:采用骨架区(framework region,FR)引物FR2、FR3和重链连接区(joining region of heavy chain,JH)引物LJH、VLJH组合、A管+B管模式、半巢式聚合酶链式反应(polymerase chain reaction,PCR)法对121例B-NHL、58例T细胞性非霍奇金淋巴瘤(T-cell non-Hodgkin lymphoma,T-NHL)和19例淋巴结反应性增生的石蜡组织进行IgH基因单克隆重排检测,分析IgH基因单克隆重排检出率在B-NHL组、T-NHL组和淋巴结反应性增生组中的差异,以及B-NHL中联合应用FR2和FR3与单独应用FR2、FR3之间IgH基因单克隆重排检出率的差异。结果:118例成功检测的B-NHL中,IgH基因单克隆重排检出率为81%(96/118);54例成功检测的T-NHL中,IgH基因单克隆重排检出率为4%(2/54);19例成功检测的淋巴结反应性增生中未检出IgH基因单克隆重排。B-NHL组与T-NHL组、淋巴结反应性增生组相比,IgH基因单克隆重排检出率差异具有显著性(P<0.05)。B-NHL中,FR2基因单克隆重排检出率为58%(68/118),FR3基因单克隆重排检出率为55%(65/118),联合应用FR2和FR3,IgH基因单克隆重排检出率为81%(96/118),联合应用FR2和FR3与单独应用FR2、FR3的检出率有显著差异(P<0.05)。结论:采用FR2、FR3、LJH及VLJH引物组合、A管+B管模式和半巢式PCR法进行石蜡组织IgH基因单克隆重排检测,简单易行,结果准确可靠,阳性率较高,可用于临床B-NHL的辅助诊断。AIM： To establish a reliable and feasible protocol for detection of monoclonal immunoglobulin heavy chain（IgH） gene rearrangements for routine diagnosis of B-cell non-Hodgkin lymphoma（B-NHL）.METHODS： Using the primer combinations of FR2,FR3,LJH and VLJH,mode tube A＋ tube B,and semi-nested PCR,the monoclonal IgH gene rearrangements in 121 cases of B-NHL,58 cases of T-cell non-Hodgkin lymphoma（T-NHL） and 19 cases of reactive lymphoid hyperplasia were detected.The differences of clonality detection rate between B-NHL group and T-NHL group,B-NHL group and reactive lymphoid hyperplasia group,and between the use of FR2,FR3 and FR2＋FR3 primers were analyzed.RESULTS： The clonality detection rates of B-NHL,T-NHL and reactive lymphoid hyperplasia were 81%（96/118）,4%（2/54） and 0%（0/19）.There were remarkable differences between B-NHL group and T-NHL group,B-NHL group and reactive lymphoid hyperplasia group in monoclonal IgH gene rearrangements（P0.05）.In B-NHL group,monoclonality was found in 58% of the cases using primer FR2,55% using FR3,and 81% using the combination of both primers,with significant differences（P0.05）.CONCLUSION： Using the primer combinations of FR2,FR3,LJH and VLJH,detection of paraffin-embedded tissues,the method of tube A＋tube B mode and semi-nested PCR for determining monoclonal IgH gene rearrangements is feasible and reliable,and the clonality detection rate is high enough for clinical diagnosis of B-NHL.

关 键 词：淋巴瘤 免疫球蛋白 基因重排 

分 类 号：R733.4[医药卫生—肿瘤]