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. 2010 Jun 6;25(3):168–176. doi: 10.1007/s12250-010-3106-4

Establishment of an ELISA to detect Kaposi’s sarcoma-associated herpesvirus using recombinant ORF73

Xin-xing Ouyang 1,3, Bi-shi Fu 1,3, Bao-lin Li 1,3, Yan Zeng 4, Fan-hong Xu 5,✉, Lin-ding Wang 1,2,✉
PMCID: PMC8227903  PMID: 20960290

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).

Key words: Kaposi’s sarcoma-associated herpesvirus (KSHV), ORF73, ELISA, Hubei, Xinjiang, Seroprevalence

Footnotes

Foundation items: Supported by the Research grants from Mega Scientific Project for HIV in China (2008ZX-10001-002), National Natural Science Foundation of Hubei Province (2008CDA013), the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and National Institutes of Health (DE017333); Open Research Fund Program of the State Key Laboratory of Virology of China (2010012).

Contributor Information

Fan-hong Xu, Phone: +86-21-62803189, Email: xfh11@21cn.com.

Lin-ding Wang, Phone: +86-27-87197600, FAX: +86-27-87197600, Email: wangld@wh.iov.cn.

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Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

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