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Detection of Antibody to hepatitis delta virus in Human Serum by Double antigen Sandwich ELISA

Virologica Sinica 2009年第1期24卷 45-51页

作者： Li XIE De-zhuang HUANG Li-xiang HE Zhao-xia LUO Yu-sen ZHOU Xiao-dong WU Beijing You-An Hospital Capital University of Medical Science Beijing 100069 China Beijing Institute ofMicrobiology and Epidemiology Beijing 100071 China Department of Biology Graduate University ofChinese Academy of Sciences Beijing 100049 China

A simple rapid detection of antibody to hepatitis delta virus (anti-HDV) in human serum was developed by using double antigen sandwich ELISA. HDV gene fragment encoding HDAg was isolated from a Chinese patient infected with HDV by RT-PCR, and a high-efficient expression HD-PQE31 strain was constructed with the fragment. We obtained high titer and good quality hepatitis delta virus protein purified by Ni-NTA metal-affinity chromatography, which was identified by Western blot and ELISA, then we set up the double antigen sandwich ELISA for detection of anti-HDV in human serum, and the performance of the sandwich ELISA was evaluated in terms of specificity and sensitivity. Results were: 1) The purified HDAg protein’s purity was 90%, and its ELISA titer was 1/100 000. 2) 42 anti-HDV positive sera were detected and showed that the sensitivity of sandwich ELISA was higher than that of competitive ELISA (t=2.44, phepatitis viruses and 30 normal samples were negative for anti-HDV. These results suggested that the double antigen sandwich ELISA with purified recombinant HDAg showed higher specificity and sensitivity, It can be used in routine laboratories to diagnose the HDV infection.

关键词： hepatitis delta virus hepatitis delta virus antigen Anti-HDV Sandwich ELISA

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