Establishment of an ELISA to detect Kaposi's sarcoma-associated herpesvirus using recombinant ORF73.
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).
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Xin-xing
XX
Ouyang
Bi-shi
BS
Fu
Bao-lin
BL
Li
Yan
Y
Zeng
Fan-hong
FH
Xu
Lin-ding
LD
Wang
D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics; D015153:Blotting, Western; D002681:China / Q000453:epidemiology; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000453:epidemiology* / Q000821:virology; D019288:Herpesvirus 8, Human / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016036:Seroepidemiologic Studies; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D014773:Virology / Q000379:methods*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins