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目的 建立小儿人巨细胞病毒 (HCMV)感染的分子诊断方法。方法 采用荧光定量聚合酶链反应 (FQ PCR)法测定 45例疑诊HCMV感染的患儿 ,并与常规PCR及酶联免疫吸附试验(ELISA)比较。对确诊为HCMV肝炎的 2 5例分别在 5个时点 (治疗前、出院时、3个月、6个月及 9个月 )监测其外周血白细胞中HCMVDNA拷贝 (copies)数。结果 常规PCR、ELISA和FQ PCR的阳性率分别为 60 0 0 % ,33 33 %和 66 67% ,灵敏度分别为 84 38% ,46 88%和 93 75 %。HCMV肝炎治疗组和对照组的HCMVDNA量在 5个时点的差异有非常显著意义 (P <0 0 0 1 )。以 1 0 3 copies ml的FQ PCR值为临界值 (≥ 1 0 3 有症状 ,<1 0 3 无症状 )可预测活动性HCMV感染的出现。结论 FQ PCR可作为小儿HCMV感染的有效诊断方法之一 ,动态检测HCMVDNA含量有助于指导治疗、估计疾病的发展和预后。
Objective To establish a molecular diagnostic method for human cytomegalovirus (HCMV) infection in children. Methods Forty-five children with suspected HCMV infection were detected by fluorescence quantitative polymerase chain reaction (FQ PCR) and compared with routine PCR and enzyme-linked immunosorbent assay (ELISA). The number of HCMVDNA copies in peripheral leukocytes was monitored at 5 time points (before treatment, at discharge, at 3 months, 6 months and 9 months) in 25 cases diagnosed as HCMV hepatitis. Results The positive rates of PCR, ELISA and FQ PCR were 60 0%, 33 33% and 66 67%, respectively. The sensitivity was 84 38%, 46 88% and 93 75% respectively. HCMV hepatitis treatment group and control group HCMVDNA at five time points the difference was significant (P <0 0 0 1). The presence of active HCMV infection was predictive of FQ PCR values of 1 0 3 copies ml as a cutoff value (≥103 symptomatic, <103 asymptomatic). Conclusion FQ PCR can be used as one of the effective diagnostic methods for HCMV infection in children. The dynamic detection of HCMVDNA content can help guide the treatment and estimate the development and prognosis of HCMV infection in children.