克隆性基因重排检测用于诊断B细胞淋巴瘤微小病灶

目的探讨B细胞淋巴瘤（B-NHL）外周血及骨髓克隆性基因重排检测对微小病灶（MD）的诊断价值。方法应用PCR技术，扩增IgH、bcl-2／IgH（包括MBR及MCR）基因重排，对42例治疗前B-NHL患者外周血及骨髓同时进行MD检测，对照组为25例非B-NHL。用IgH基因重排阳性的淋巴瘤CA46细胞株作系列稀释法实验，判断以IgH为分子标志检测外周血及骨髓MD实验的灵敏度。结果淋巴瘤CA46细胞株系列稀释法实验显示：以IgH基因重排为分子标志检测MD敏感性为10^-2 。42例B-NHL外周血中IgH检出率19％（8／42），bcl-2／IgH检出率为33．3％（14／42）。对42例B-NHL同时进行的骨髓MD检测中，IgH检出率为28．6％（12／42），bcl-2／IgH MBR检出率为45．2％（19／42）。外周血及骨髓中T细胞淋巴瘤及其他淋巴瘤3种指标检测均为阴性。IgH、bcl-2／IgH MBR、bcl-2／IgH MCR3个指标各自在骨髓中检出率与外周血中检出率差别无统计学意义（P〉O．05），虽然bcl-2／IgH MBR检出率在外周血与骨髓中滤泡性淋巴瘤（FCL）组均较弥漫性大B细胞淋巴瘤组高，但差别无统计学意义（P〉0．05）。在同时进行的外周血及骨髓检测中，12例骨髓IgH阳性中有8例外周血检测亦为阳性，19例骨髓bcl-2／IgH MBR阳性中有13例外周血检测为阳性，但1例FCL骨髓检测阴性外周血检测为阳性。结论检测外周血bcl-2／IgH MBR可作为诊断B-NHL MD辅助手段。

Clonal Gene Rearrangement for the Detection of Minimal Residual Lesions in B Cell Lymphoma

Objective To analyze the value of clonal gene rearrangement for minimal disease (MD) in peripheral blood (PB) and bone marrow (BM) of the patients with B cell lymphoma (B-NHL). Methods semi-nested/nested polymerase chain reaction were used to amplify the IgH and bcl-2/IgH gene rearrangement in the specimens of B-NHL, PB and BM specimens from 42 untreated B-NHL and 25 untreated non-B-NHL for detection of minimal nesidual lesions. Serial dilution test of the lymphoma cell line CA46 was done to study the sensitivity of detection MD with IgH as a marker. Results The lymphoma cell line CA46 serial dilution test showed the sensitivity of detection MD with the gene rearrangement of IgH was 10 -2. In peripheral blood , IgH was positive in 8 out of 42 B-NHL specimens, the positive rate was 19%(8/42), and 20%(6/30) and 16.7%(2/12) in diffuse large B cell lymphoma(DLBCL) and follicular cell lymphoma(FCL) specimens respectively. The positive rate of bcl-2/IgH was 33.3%(14/42), and 30%(9/30) and 41.7%(5/12) in DLBCL and FCL specimens respectively. In BM, the positive rate of IgH was 28.6%(12/42), and 33.3%(10/30) and 16.7%(2/12) in DLBCL and FCL specimens respectively. The positive rate of bcl-2/IgH was 45.23%(19/42), and 36.7%(11/30) and 66.7%(8/12) in DLBCL and FCL specimens respectively. All bcl-2/IgH in positive specimens belong to bcl-2/IgH,MBR no bcl-2/IgH MCR. None of positive was showed in 25 non-B-NHL cases, neither in PB nor in BM. There was no signification between PB and BM of positive rate of IgH, bcl-2/IgH MBR and bcl-2/IgH MCR (P>0.05). Although the bcl-2/IgH MBR positive rate of FCL was higher than that of DLBCL(both in PB and in BM), there was no signification between them(P>0.05). Conclusion The detection of bcl-2/IgH MBR in PB may be a useful ancillary method to detect minimal residual in B-NHL.