论文部分内容阅读
巨细胞病毒感染的血清学诊断方法是目前临床诊断中应用最广泛的方法之一。但由于缺少高效价、高纯度和高特异性的抗原，使各种血清学方法在其敏感性和特异性方面均不能令人满意。文献报道，通过分子生物学得到的或化学合成的单一病毒蛋白或部分成份可作为血清学诊断的良好抗原。本文报道用聚合酶链反应扩增了巨细胞病毒主要磷蛋白ｐＰＵＬ３２的一个强抗原决定簇基因，并同时在扩增产物的两端引入了相应的限制性内切酶识别位点序列，经与表达载体连接后转入相应的细菌中，得到了能够表达此抗原决定簇的克隆，其表达的融合蛋白在免疫转印检测中与人巨细胞病毒阳性血清有强特异性反应。
Serological diagnosis of cytomegalovirus infection is one of the most widely used clinical diagnostic methods. However, due to the lack of high titer, high purity and high specificity of antigens, various serological methods are not satisfactory in terms of their sensitivity and specificity. It has been reported in the literature that single virus proteins or partial components obtained by molecular biology or chemically synthesized can be good antigens for serodiagnosis. In this paper, a strong antigenic determinant gene of cytomegalovirus major phosphoprotein pPUL32 was amplified by polymerase chain reaction (PCR). At the same time, a restriction enzyme recognition site sequence was introduced at both ends of the amplified product. After the expression vector was ligated and transferred into the corresponding bacteria, a clone capable of expressing the antigenic determinant was obtained. The expressed fusion protein strongly reacted with human cytomegalovirus positive sera in the immunoblot assay.