巨细胞病毒gp52蛋白表达与应用  

Cloning and application of CMV gp52 protein

作　　者：吴丽霞[1] 赵玉红[1] 陈咏君[1] 郭晶[1] 孙超[1] 张小刚 于庭[3] 李凤娣[4] 杨丽荣[5] 

机构地区：[1]沈阳医学院沈洲医院检验科,沈阳110002 [2]北京现代高达生物技术有限责任公司 [3]吉林大学第二医院 [4]沈阳市疾病预防控制中心 [5]沈阳市大东区妇幼保健所

出　　处：《中国公共卫生》2008年第12期1495-1497,共3页Chinese Journal of Public Health

摘　　要：目的构建表达巨细胞病毒(CMV)gp52蛋白抗原的重组质粒及工程菌,获取纯化的gp52蛋白抗原。方法采用PCR技术,克隆表达CMVgp52重组蛋白,利用产物建立IgM捕获ELISA方法鉴定其抗原性及实用性。结果表达纯化的gp52蛋白纯度>95%,经酶标记建立IgM捕获ELISA方法,检测35份抗巨细胞病毒IgM阳性血清和35份阴性血清,用酶标记gp52蛋白建立的捕获ELISA法阳性检出率97.1%,阴性检出率100%,与意大利SORIN公司试剂盒检测结果比较,差异无统计学意义(P>0.05);其中1份CMV-IgM阳性血清1:16稀释后仍能与抗原反应,表明gp52蛋白抗原表位有较好的抗原特异性。结论高效表达纯化的gp52蛋白抗原性强,利用其建立的IgM捕获ELISA方法,可用于检测巨细胞病毒抗体。Objective To construct the recombinant plasmid and engineering bacteria for expression of CMV gp52 protein and to use the expressed product for detection of CMV specific antibody. Methods Using PCR technology, recombinant CMV gp52 was cloned and expressed to establish IgM capture ELISA, and the product was detected for its antigenicity and practicality. Results Through expression and purification, the purity of gp52 was over 95 96. IgM capture ELISA established by gp52 - HRP was used to detect 35 CMV - positive serums and 35 CMV- negative serums, the positive detection rate was 97.1% and negative detection rate was 100 96 by capture ELISA with no significant difference compared with SORIN company＇ kit（P 〉 0.05）. gp52 antigen can response to the one CMV - positive serum （ 1 ： 16 dilution）, so the gp52 had better antigenicity. Conclusion CMV gp52 protein efficiently expressed had specific antigenicity, and can be used for establishing IgM capture ELISA to detect cytomegalovirus antibodies.

关 键 词：巨细胞病毒 gp52蛋白 抗原 基因克隆 

分 类 号：R373[医药卫生—病原生物学]