IgH基因重排和免疫分型检测在诊断B淋巴细胞增生性疾病中的初步应用

Diagnosis of B lymphocyte proliferative disorders by the detection of IgH gene rearrangement and immunologicaltype

目的通过半巢式聚合酶链反应(PCR)技术对免疫球蛋白重链(IgH)基因重排的检测,建立对B淋巴细胞增生性疾病早期诊断的分子生物学方法,进而探讨在疑似B淋巴细胞增生性疾病诊断与鉴别诊断中的价值。方法通过半巢式PCR技术检测8例已确诊的B淋巴细胞增生性疾病和24例临床疑似B淋巴细胞增生性疾病初诊患者的IgH基因重排;24例中流式细胞术检测了19例患者的免疫分型。结果检测8例已确诊的B淋巴细胞增生性疾病,该方法敏感性为83.33%(5/6),特异性为100%(10/10);24例疑似B淋巴细胞增生性疾病初诊患者IgH基因重排检出的阳性率为83.33%(20/24),其中FR2A阳性率41.67%(10/24);FR3A阳性率50%(12/24);同时阳性2例。免疫分型结果14例可疑阳性,阳性率73.68%(14/19)。结论通过建立半巢式聚合酶链式反应(Polymerase chain reaction,PCR)技术检测IgH基因重排的分子生物学方法,对疑似B淋巴细胞增生性疾病患者的诊断起到了十分重要的作用,并且对反应性和肿瘤性淋巴细胞增生鉴别诊断具有重要的意义,为临床早期诊断、治疗干预提供了新的证据。

objective To establish the semi - nested PCR technology to detect immunoglobulin heavy chain （IgH） gene rearrangement, in diagnosis of early B lymphocyte proliferative disorders, and appraise its value in diagnosis and differential diagnosis of doubtful B lymphocyte proliferative disorders. Methods Eight patients with definite and 24 patients with doubtful B lymphocyte proliferative disorders were investigated using semi - PCR technology; and immunologicaltype was assayed in 19 of the 24 patients by flow cytometry. Results In the 8 patients with definite diagnosis of B lymphocyte proliferative disorders,the sensitivity of PCR was 83.33% （5/6） and the specificity was 100%. In the 24 patients with suspected B lymphocyte proliferative disorders, the positive rate of IgH gene rearrangement was 83.33% （20/24）, FR2A positive rate 41.67% （10/24）, and FR3A positive rate 50% （12/24）. Fourteen cases were doubtfully positive to detection of immunologicaltype, the positive rate being 73.68% （14/19）.Conclusions A semi - nested PCR technology has been established for detecting IgH gene rearrangement, which plays an important role in the diagnosis of doubtful B lymphocyte proliferative disorders, and is of significance in differential diagnosis of reactive and tumour lymphocyte proliferation, and provides a new evidence for early diagnosis and therapy in clinic.