FQ-PCR检测造血干细胞移植受者HCMV感染及与淋巴细胞免疫表型变化的关系

Detection of cytomegalovirus infection by FQ-PCR in patients after allogeneic peripheral blood stem cell transplantation and its relation with changes of peripheral lymphocyte subsets

目的探讨实时荧光定量聚合酶链反应（FQ-PCR）检测人巨细胞病毒（HCMV）DNA含量在异基因外周血造血干细胞移植（allo-PBSCT）受者HCMV激活感染中的诊断价值和淋巴细胞免疫表型变化与HCMV激活感染的关系。方法应用FQ-PCR和酶联免疫吸附试验（ELISA）检测allo-PBSCT后受者HCMV DNA和抗HC-MV lgM抗体;用流式细胞仪测定淋巴细胞亚群。结果39例allo-PBSCT受者中19例检出HCMV DNA（〈30 d的有5例,2-3个月的13例,〉100 d的1例）。HCMV感染后血清HCMV、IgM阳性检出率非常低（4/19）,且出现时间较晚。移植后早期HCMV DNA阳性组与阴性组及健康组比较CD4^＋细胞、CD19^＋细胞及CD4/CD8比值显著下降,CD8^＋细胞和CD16＋56^＋细胞升高。HCMV阳性患者CD4/CD8比值在感染前后比较差异也有统计学意义。结论allo-PBSCT后HCMV活动性感染易发生于allo-PBSCT后早、中期,高峰期在移植后30-60 d;HC-MV活动性感染与细胞免疫功能紊乱互为因果,加重病情。

Objective To explore the usefulness of fluorescence quantitative polymerase chain reaction（FQ-PCR） for early detection of human cytomegalovirus（HCMV）DNA,and to investigate the relation between changes of peripheral lymphocyte subsets and the influence of active HCMV infection on immunity in patients after allogeneic peripheral blood stem cell transplantation（allo-PBSCT）. Methods HCMV DNA and HCMV IgM were detected by FQ-PCR and enzyme-linked immunosorbent assay（ELISA） respectively.Peripheral lymphocyte subsets were assayed by flow cytometry. Results In 39 patients after allo-PBSCT,19 patients were HCMV DNA positive（5 cases≤30 days,13 cases 2-3 month,1 case〉100 days）.HCMV IgM antibody was detected in the late stage after transplantation and the positive rate was very low（4/19）.In the early stage after allo-PBSCT,the percentage of CD4^＋,CD19^＋and CD4/CD8 significantly decreased in patients with positive HCMV DNA compared with HCMV DNA negative group and healthy control.While,the level of CD8^＋ and CD16＋56^＋ increased in infection group.There was statistic difference in CD4/CD8 before and after HCMV infection. Conclusions Active HCMV infection appeared frequently in the early or middle stage of allo-PBSCT,especially 30-60 days.Active HCMV infection and the abnormal immune function influence mutually,leading to the exacerbation of disease.