摘要
目的进一步分析优化结核杆菌抗原优势表位,获得高特异性融合抗原,以满足研制结核病检测试剂的需要。方法用分子生物学软件分析结核杆菌四种抗原MPT64、Mtb8、Mtb8.4和Mtb16.3的优势表位后,分别插入到pBVIL1载体中,构建表达质粒,并利用pBVIL1载体特点,将其基因连接后,进行融合表达,经纯化获得抗原,用间接ELISA法测其活性。结果各抗原优势肽段及融合抗原均获得了高效表达,融合抗原的特异性和灵敏性均高于各单一肽段。结论此融合抗原可用于研制结核病的诊断试剂。
Objective In order to obtain the fused antigen with high specificity and high sensitivity which was used to detect the tuberculosis.Methods The dominant epitopes of four antigens of Mycobacterium tuberculosis,MPT64,Mtb8,Mth8.4 and Mtbl6.3, were analyzed using the BIOSUN software.The gene sequence of each dominant antigen peptide was cloned by PCR,and then ligated into expression vector pBVIL1. The genes were induced to express in E coli HB101 and the anfigenicity of each antigen was detected by indirect ELISA. Results Each dominant antigen peptide and the fused antigen were expressed highly in E coli HB101. The fused antigen presented more favourable antigenicity than that of each dominant antigen peptide. Conclusion The fused antigen, 8.4- 64 - 16.3- 8,might serve as a serediagnostic candidate antigen and was used to detect the tubeulosis,
出处
《中国实验诊断学》
2008年第5期569-573,共5页
Chinese Journal of Laboratory Diagnosis
基金
国家863重大专项2006AA02090804
国家自然科学基金30772067
