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Expression of Kaposi’s sarcoma-associated herpesvirus ORFK8.1 and its preliminary diagnostic application

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Virologica Sinica

Abstract

The ORFK8.1 of Kaposi’s sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogalactopyranoside (IPTG). The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis (SDS-PAGE). SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant ORFK8.1 protein. The optimal condition of the recombinant ORFK8.1 ELISA assay was confirmed: the concentration of antigen was 5 μg/mL, the dilution of serum was 1:200. We used the ELISA method to investigate the recombinant ORF K8.1 protein’s specificity, the data showed that the specificity of ORF K8.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFK8.1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.

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Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China

    Bi-shi Fu, Bao-lin Li, Xin-xing Ouyang & Lin-ding Wang

  2. Graduate School of the Chinese Academy of Sciences, Beijing, 100039, China

    Bi-shi Fu, Bao-lin Li & Xin-xing Ouyang

  3. Shanghai Institute of Biological Products, Shanghai, 200052, China

    Fan-hong Xu

  4. Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University, Shihezi, 832002, China

    Yan Zeng

Authors
  1. Bi-shi Fu
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  2. Bao-lin Li
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  3. Xin-xing Ouyang
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  4. Yan Zeng
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  5. Fan-hong Xu
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  6. Lin-ding Wang
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Correspondence to Fan-hong Xu or Lin-ding Wang.

Additional information

Foundation items: Supported by the Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences (0702121YJ1); Open Research Fund Program of the State Key Laboratory of Virology of China (2007013).

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Fu, Bs., Li, Bl., Ouyang, Xx. et al. Expression of Kaposi’s sarcoma-associated herpesvirus ORFK8.1 and its preliminary diagnostic application. Virol. Sin. 24, 202–208 (2009). https://doi.org/10.1007/s12250-009-3029-0

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  • DOI: https://doi.org/10.1007/s12250-009-3029-0

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