Abstract
目的 用我们研制的一种新人巨细胞病毒(HCMV)重组被膜磷蛋白pp65为抗原,建立快速、简便和特异的检测HCMV IgM 的间接酶联免疫吸附法(REC-ELISA).方法用自行设计、表达的新的HCMV pp65重组抗原,建立REC-ELISA,并检测100份临床疑似为HCMV感染者血清中HCMV-IgM,同时与全病毒为抗原的间接ELISA法(简称全病毒-ELISA法)和美国BioCheck试剂盒(简称BioCheck法)检测结果进行比较.结果 REC- ELISA法重组抗原最适量为3.5 μg/孔,HRP标记二抗为1∶ 800,血清稀释度为1∶ 100; 稳定性试验阳性变异系数(CV)为9.5%,重复性试验平均CV值:批内CV 4.5%,批间CV 9.6%.REC-ELISA法、全病毒-ELISA法和BioCheck法检测HCMV IgM的阳性率分别为44%、50%和45%;REC-ELISA法的敏感性为95.6%,其特异性(98.2%)高于全病毒-ELISA法(90.9%);正确指数为92.8%,与BioCheck法一致性为97.0%.结论用pp65重组抗原建立的REC-ELISA法具有高度特异性与敏感性,且操作简单、快速,重复性好;与全病毒抗原相比,重组抗原可规模化、标准化生产,且更安全、经济,具有良好的临床应用前景。
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