Citation: OUYANG Xin-xing, FU Bi-shi, LI Bao-lin, ZENG Yan, XU Fan-hong, WANG Lin-ding. Establishment of an ELISA to Detect Kaposios Sarcoma-associated Herpesvirus Using Recombinant ORF73 .VIROLOGICA SINICA, 2010, 25(3) : 168-176.  http://dx.doi.org/10.1007/s12250-010-3106-4

Establishment of an ELISA to Detect Kaposios Sarcoma-associated Herpesvirus Using Recombinant ORF73

  • Corresponding author: XU Fan-hong, xfh11@21cn.com
    WANG Lin-ding, wangld@wh.iov.cn
  • Received Date: 23 October 2009
    Accepted Date: 31 March 2010
    Available online: 01 June 2010

    Fund Project: Open Research Fund Program of the State Key Laboratory of Virology of China 2010012the Research grants from Mega Scientific Project for HIV in China 2008ZX- 10001-002National Natural Science Foundation of Hubei Province 2008CDA013the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and National Institutes of Health DE017333

  • Kaposi’s sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi’s sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman’s disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Han people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).

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    Establishment of an ELISA to Detect Kaposios Sarcoma-associated Herpesvirus Using Recombinant ORF73

      Corresponding author: XU Fan-hong, xfh11@21cn.com
      Corresponding author: WANG Lin-ding, wangld@wh.iov.cn
    • 1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
    • 2. Department of Microbiology, Anhui Medical University, Hefei 230032, China
    • 3. Graduate School of the Chinese Academy of Sciences, Beijing 100039, China
    • 4. Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University, Shihezi 832002, China
    • 5. Shanghai Institute of Biological Products, Shanghai 200052, China
    Fund Project:  Open Research Fund Program of the State Key Laboratory of Virology of China 2010012the Research grants from Mega Scientific Project for HIV in China 2008ZX- 10001-002National Natural Science Foundation of Hubei Province 2008CDA013the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and National Institutes of Health DE017333

    Abstract: Kaposi’s sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi’s sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman’s disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Han people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).