重组人巨细胞病毒(HCMV)优势表位嵌合抗原的构建表达以及捕获法检测IgM抗体

Capture and detection of human cytomegalovirus (HCMV) IgM by a recombinant multi-epitope chimeric antigen

通过计算机软件分析选择巨细胞病毒优势抗原表位,用聚合酶链式反应(PCR)扩增优势表位基因,构建了巨细胞病毒多优势表位嵌合抗原表达载体p4T-UL32-UL44-UL80a-UL83,转化宿主菌大肠杆菌BL21(DE3)进行表达,GST亲和层析法获得高纯度嵌合抗原,辣根过氧化物酶(HRP)标记嵌合抗原,并构建捕获法检测血清中的抗HCMVIgM抗体。结果表达的优势表位嵌合抗原具有很好的抗原性,用其建立的捕获法检测确诊的30份阳性和63份儿童阴性血,阳性检出率和阴性检出率都是100%说明用嵌合抗原构建的捕获法检测血清中抗HCMVIgM抗体具有更高的灵敏度和检出正确率,其检测水平已经达到国外的同类产品水平。

To establish a sensitive and specific antibody-capture enzyme-linked immunosorbent assay （AC-ELISA） method to detect serum IgM against human cytomegalovirus （HCMV） by expressing a recombinant HCMV multi-epitope cheimeric antigen through genetic engineering. The dominant epitopes of HCMV were analyzed and selected by computer software A recombinant multi-epitope chimeric antigen expression vector including HCMV DNA was constructed, and transformed into E. coli BL21（DE3）. The antigen was abundantly expressed, purified, and labeled by horseradish peroxidase for subsequent development of the AC-ELISA. Thirty validated positive sera and sixty-three validated negative sera were submitted for IgM detection by this recombinant antigen. The sensitivity and specificity of AC-ELISA with our recombinant antigen were both 100%. The sensitivity and specificity of this AC-ELISA diagnostic kit with the recombinant antigen are comparable to similar foreign commercial products.