Serving the Society Since 1986
Advanced Search
Volume 24 Issue 3
June 2009
    Citation: Bi-shi FU, Bao-lin LI, Xin-xing OUYANG, Yan ZENG, Fan-hong XU, Lin-ding WANG. Expression of Kaposi's Sarcoma-associated Herpesvirus ORFK8.1 and Its Preliminary Diagnostic Application* .VIROLOGICA SINICA, 2009, 24(3) : 202-208.  http://dx.doi.org/10.1007/s12250-009-3029-0
    shu

    Expression of Kaposi's Sarcoma-associated Herpesvirus ORFK8.1 and Its Preliminary Diagnostic Application*

    cstr: 32224.14.s12250-009-3029-0
    • 1.

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

    • 2.

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

    • 3.

      Shanghai Institute of Biological Products, Shanghai 200052, China

    • 4.

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

    • Corresponding author: Fan-hong XU, xfh11@21cn.com
      Lin-ding WANG, wangld@wh.iov.cn
    • Received Date: 14 January 2009
      Accepted Date: 14 April 2009
      Available online: 01 June 2009

      Fund Project: Open Research Fund Program of the State Key Laboratory of Virology of China 2007013the Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences 0702121YJ1

    • 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%.
    • 加载中

      1. Albrecht J C, Nicholas J, Cameron K R, et al. 1992. Herpesvirus saimiri has a gene specifying a homologue of the cellular membrane glycoprotein CD59. Virology, 190: 527-530.
          doi: 10.1016/0042-6822(92)91247-R

      2. Baillargeon J, Deng J H, Hettler E, et al. 2001. Sero-prevalence of Kaposi's sarcoma-associated herpesvirus infection among blood donors from Texas. Ann Epidemiol, 11: 512-518.
          doi: 10.1016/S1047-2797(01)00242-3

      3. Belec L, Cancre N, Hallouin M C, et al. 1998. High prevalence in Central Africa of blood donors who are potentially infectious for human herpesvirus 8. Transfusion, 38: 771-775.
          doi: 10.1046/j.1537-2995.1998.38898375517.x

      4. Cesarman E, Chang Y, Moore P S, et al. 1995. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. N Engl J Med, 332: 1186-1191.
          doi: 10.1056/NEJM199505043321802

      5. Chang Y, Cesarman E, Pessin M S, et al. 1994. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science, 266: 1865-1869.
          doi: 10.1126/science.7997879

      6. Engels E A, Sinclair M D, Biggar R J, et al. 2000. Latent class analysis of human herpesvirus 8 assay performance and infection prevalence in sub-saharan Africa and Malta. Int J Cancer, 88: 1003-1008.
          doi: 10.1002/(ISSN)1097-0215

      7. Engels E A, Whitby D, Goebel P B, et al. 2000. Identifying human herpesvirus 8 infection: performance characteristics of serologic assays. J Acquir Immune Defic Syndr, 23: 346-354.
          doi: 10.1097/00042560-200004010-00011

      8. Fang Q, Liu J, Bai Z Q, et al. 2006. Seroprevalence of Kaposi's sarcoma-associated herpesvirus in the central population from Hubei Province. Virologica sinica, 21: 97-101.

      9. Fujii T, Taguchi H, Katano H, et al. 1999. Seroprevalence of human herpesvirus 8 in human immunodeficiency virus 1-positive and human immunodeficiency virus 1-negative populations in Japan. J Med Virol, 57:159-162.
          doi: 10.1002/(ISSN)1096-9071

      10. Gao S J, Kingsley L, Li M, et al. 1996. KSHV antibodies among Americans, Italians and Ugandans with and without Kaposi's sarcoma. Nat Med, 2: 925-928.
          doi: 10.1038/nm0896-925

      11. Lomonte P, Bublot M, van Santen V, et al. 1995. Analysis of bovine herpesvirus 4 genomic regions located outside the conserved gammaherpesvirus gene blocks. J Gen Virol, 76 (Pt 7): 1835-1841.

      12. Melbye M, Cook P M, Hjalgrim H, et al. 1998. Risk factors for Kaposi's-sarcoma-associated herpesvirus (KSHV/ HHV-8) seropositivity in a cohort of homosexual men, 1981-1996. Int J Cancer, 77: 543-548.
          doi: 10.1002/(ISSN)1097-0215

      13. Raab M S, Albrecht J C, Birkmann A, et al. 1998. The immunogenic glycoprotein gp35-37 of human herpesvirus 8 is encoded by open reading frame K8.1. J Virol, 72: 6725-6731.

      14. Russo J J, Bohenzky R A, Chien M C, et al. 1996. Nucleotide sequence of the Kaposi sarcoma-associated herpesvirus (HHV8). Proc Natl Acad Sci USA, 93: 14862-14867.
          doi: 10.1073/pnas.93.25.14862

      15. Schulz T F. 1998. Kaposi's sarcoma-associated herpes-virus (human herpesvirus-8). J Gen Virol, 79 (Pt 7): 1573-1591.

      16. Simpson G R, Schulz T F, Whitby D, et al. 1996. Prevalence of Kaposi's sarcoma associated herpesvirus infection measured by antibodies to recombinant capsid protein and latent immunofluorescence antigen. Lancet, 348: 1133-1138.
          doi: 10.1016/S0140-6736(96)07560-5

      17. Soulier J, Grollet L, Oksenhendler E, et al. 1995. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease. Blood, 86: 1276-1280.

      18. Virgin H W t, Latreille P, Wamsley P, et al. 1997. Complete sequence and genomic analysis of murine gammaherpesvirus 68. J Virol, 71: 5894-5904.

      19. Whitby D, Luppi M, Barozzi P, et al. 1998. Human herpesvirus 8 seroprevalence in blood donors and lymphoma patients from different regions of Italy. J Natl Cancer Inst, 90: 395-397.
          doi: 10.1093/jnci/90.5.395

    • 加载中

    Figures(3) / Tables(1)

    PDF

    Article Metrics

    Article views(4371) PDF downloads(15) Cited by()

    Related
    Proportional views

      Expression of Kaposi's Sarcoma-associated Herpesvirus ORFK8.1 and Its Preliminary Diagnostic Application*

        Corresponding author: Fan-hong XU, xfh11@21cn.com
        Corresponding author: Lin-ding WANG, wangld@wh.iov.cn
      • 1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
      • 2. Graduate School of the Chinese Academy of Sciences, Beijing 100039, China
      • 3. Shanghai Institute of Biological Products, Shanghai 200052, China
      • 4. Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University, Shihezi 832002, China
      • Received Date: 14 January 2009
      • Accepted Date: 14 April 2009
      Fund Project:  Open Research Fund Program of the State Key Laboratory of Virology of China 2007013the Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences 0702121YJ1

      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%.

        HTML