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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 recombinant plasmids and engineered bacteria expressing gp52 antigen of cytomegalovirus (CMV) and obtain purified gp52 protein antigen. Methods The recombinant protein of CMVgp52 was cloned and expressed by PCR. The antigenicity and practicability of the recombinant protein were identified by ELISA. Results The purity of gp52 protein was> 95%. The IgM-capture ELISA was established by enzyme-linked immunosorbent assay (ELISA). 35 positive anti-CMV sera and 35 negative sera were detected. 97.1%, negative detection rate of 100%, with the Italian company SORIN kit test results, the difference was not statistically significant (P> 0.05); one of CMV-IgM positive serum diluted with 1:16 still react with the antigen, Gp52 protein epitopes showed good antigen specificity. Conclusion The highly expressed and purified gp52 protein has strong antigenicity and can be used to detect cytomegalovirus antibody by using its established IgM capture ELISA.