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.pdfVIP

HEREDITAS (Beijing) 2007 11 , 29(11): 1351―1356 ISSN 0253-9772 研究报告 DOI: 10.1360/yc-007-1351 (HCMV) IgM 1 2 2 2 2 2 2 2 2 2 孙鹏 , 孙兴宝 , 胡鹏 , 田晓红 , 邓波 , 夏良雨 , 王宁燕 , 王红 , 李泓彦 , 何志强 1. , 518054; 2. , 518054 : 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 HCMV IgM 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 HCMV IgM 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。 : 人巨细胞病毒; 优势表位嵌合抗原; 捕获酶联免疫吸附法; IgM 抗体 Capture and detection of human cytomegalovirus (HCMV) IgM by a recombinant multi-epitope chimeric antigen 1 2 2 2 2 2 SUN Peng , SUN Xing-Bao , HU Peng , TIAN Xiao-Hong , DENG Bo , XIA Liang-Yu , 2 2 2 2 WANG Ning-Yan , WANG Hong , LI Hong-Yan , HE Zhi-Qiang 1. The Sixth People ’s Hospital of Shenzhen, Shenzhen 518054, China; 2. Fapon Biotech Inc., Shenzhen 518054, China Abstract: 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