巨细胞病毒早期抗原pp65和晚期抗原pp67-mRNA在移植术后活动性巨细胞病毒感染诊断中的价值

目的：通过对器官移植术后受者外周血中巨细胞病毒（CMV）早期抗原pp65和晚期抗原pp67-mRNA的监测，评价其对于诊断器官移植术后CMV感染、发病及预后的价值，为临床准确有效地分析理解检测结果提供依据。方法：收集28例器官移植受者EDTA抗凝外周血标本共120份，用免疫荧光技术（IFA）检测其中CMV早期抗原pp65；用核酸基础序列扩增法（NASBA）测定晚期CMVpp67-mRNA，绘制2组用于诊断CMV的受试者特征曲线（ROC曲线），比较2组曲线下面积。结果1120份样本中，42份为CMVpp65阳性（≥1个阳性细胞／2×105白细胞），23份为CMVpp67-mRNA阳性。28例器官移植受者中，9例CMVpp65和pp67-mRNA均为阳性（其中3例pp65和pp67-mRNA在同一时间为阳性），8例pp65为阳性而pp67-mRNA为阴性，2例pp65为阴性而pp67-mRNA为阳性。临床诊断感染CMV的患者4例，在感染早期无临床症状病毒潜伏期间，受试者工作特征（ROC）曲线下面积（AUC）pp65为0．9542，pp67-mRNA为0．6611，即pp65检测的灵敏度和特异性高于pp67-mRNA；而在出现临床症状并治疗后期，pp65的AUC为0．8300，pp67-mRNA为0．9232，pp67-mRNA的灵敏度和特异性高于pp65。根据ROC曲线查得，以每2x105白细胞中有10个阳性细胞为诊断CMV活动性感染的最佳临界值。结论：AUC结果表明，pp65、pp67-mRNA均具有诊断意义。pp65检测更适于早期CMV活动性诊断，对于提示临床开始抗病毒治疗具有早期快速的意义；pp67-mRNA检测快速、结果准确，可作为结束抗病毒治疗的参考指标；两者联合检测用于监测CMV，对于辅助临床诊断有很好的价值。

Clinical Significance of Human Cytomegalovirus pp65 Antigenemia and pp67-mRNA for Diagnosing Active Cytomegalovirus Infection in Transplant Recipients

Objective： To evaluate the clinical significance of pp65 antigen and pp67-mRNA for diagnosing human cytomegalovirus（CMV） infection and predicting prognosis in transplant recipients by monitoring both of them in the peripheral blood. By discussing the lab diadynamic criteria of detecting HCMV reactive infection, to provide evidence to help clinicians to apprehend the assay results correctly and effectively. Methods： 120 samples of peripheral blood treated with EDTA were collected from 28 patients after organ transplantation. The pp65 antigen was detected by assay（IFA） and the expression of pp67-mRNA was detected by nucleic acid sequence-based amplification（NASBA）. The areas under curive（AUC） which was designed from receiver operating characteristic（ROC） for dignosing CMV disease in two groups were compared. Results： In 28 transplant recipients, 3 samples were captured in the same patient at least. In 120 continuous blood samples coming from 28 organ recipients, 23 specimens were positive of pp67-mRNA, 42 specimens were positive（≥1 cell/2×10^5 WBC） of the pp65 antigen. In 28 organ recipients, the pp65 antigen and the expression of pp67-mRNA in 9 patients（both the pp65 antigen and the expression of pp67-mRNA were positive at the same time in 3 patients） were positive. The pp65 antigen of 8 patients was positive while the expression of pp67-mRNA was negative. The expression of pp67-mRNA of 2 patients was positive while the pp65 antigen was negative. There were 4 patients who were diagnosed the illness as clinical CMV disease. In early period of CMV infection, AUC of the pp65 ROC was 0.9542 and AUC of the pp67-mRNA ROC was 0.6611. But in the later period of therapies, AUC of the pp65 ROC was 0.8300 and AUC of the pp67-mRNA ROC was 0.9232. The critical value（cut-off） of diagnosing CMV reactive infection was 10 positive cells/2×10^5 WBC according to ROC. Conclusion： Both AUC of the pp65 and of the pp67-mRNA are of great value in diagnosis. The pp65 antigenemia assay is more su