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

 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 heavy chain，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，J_H）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。

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

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 (P<0.05). 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 (P<0.05). 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.