催化信号扩增法检测人巨细胞病毒被膜磷蛋白65的实验分析

Evaluation of catalyzed signal amplification in detecting human cytomegalovirus phosphoprotein 65

目的建立一种新型的快速诊断肾移植受者人巨细胞病毒(HCMV)活动性感染的方法。方法采用免疫组织化学催化信号扩增法,通过抗HCMV被膜磷蛋白65(pp65)单克隆抗体AAC10对外周血WBC中的HCMV pp65抗原进行标记识别。并与荧光定量PCR(FQ-PCR)方法进行比较。结果检测的102例肾移植受者中,血浆HCMV DNA阳性66例,HCMV pp65抗原阳性56例,血浆HCMV DNA阴性患者中无pp65阳性,两种方法检测的符合率为90.2%,相关性好(r= 0.831);HCMV病患者31例,两者均阳性;同时检测40名健康者,结果pp65抗原全为阴性。结论催化信号扩增法敏感、简便,能更准确地反映HCMV感染的活动状况,有助于肾移植术后HCMV病的早期诊断,并可指导抗病毒治疗。

Objective To establish a new method for rapid diagnosis of the active human cyto- megalovirus（HCMV） infection in renal transplant recipients. Methods HCMV phosphoprotein（pp） 65 antigen in peripheral blood leucocytes was detected by immunohistochemistry method and catalyzed signal amplification with HCMV monochonal antibody AAC10, and the results were compared with those of fluorescent quantitative polymerase chain reaction（FQ-PCR） assay. Results Of 102 renal transplant recipients, 66 were positive for HCMV DNA in plasma. Among them, 56 were positive for pp65. No patients whose HCMV DNA levels in plasma was negative were pp65 positive. The coincidence of two assays was 90. 2%, with high relationship（r = 0. 831）. Both HCMV DNA and pp65 were positive in 31 cases of HCMV infection. And HCMV pp65 was all negative in 40 healthy controls. Conclusions Catalyzed signal amplification is sensitive, convenient and reliable in the diagnosis of HCMV infection. This method is effective in early diagnosis of HCMV infection in renal transplant recipients and is helpful for the antiviral treatment.