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Virologica Sinica

, Volume 24, Issue 1, pp 45–51 | Cite as

Detection of antibody to hepatitis delta virus in human serum by double antigen sandwich ELISA

  • Li XieEmail author
  • De-zhuang Huang
  • Li-xiang He
  • Zhao-xia Luo
  • Yu-sen Zhou
  • Xiao-dong Wu
Article
  • 95 Downloads

Abstract

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, p<0.01). 3) The inhibitory rates for 2 anti-HDV positive sera by the specific HDAg were 74% and 93% respectively. 4) For the assay of specificity, all 60 samples infected by other hepatitis 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.

key words

Hepatitis delta virus Hepatitis delta virus antigen Anti-HDV Sandwich ELISA 

CLC number

R512.6 

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Copyright information

© Wuhan Institute of Virology, CAS and Springer-Verlag GmbH 2009

Authors and Affiliations

  • Li Xie
    • 1
    • 3
    Email author
  • De-zhuang Huang
    • 1
  • Li-xiang He
    • 1
  • Zhao-xia Luo
    • 1
  • Yu-sen Zhou
    • 2
  • Xiao-dong Wu
    • 3
  1. 1.Beijing You-An HospitalCapital University of Medical ScienceBeijingChina
  2. 2.Beijing Institute of Microbiology and EpidemiologyBeijingChina
  3. 3.Department of BiologyGraduate University of Chinese Academy of SciencesBeijingChina

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