Expression of recombinant VP1 protein of enterovirus 71 and development of serological assay for detection of EV71 infection.
- Author:
Bo-ping ZHOU
1
;
Wei-long LIU
;
Jing-jing XIE
;
Xin-chun CHEN
;
Liu-mei XU
;
Yan TAN
;
Ying-xia LIU
;
Gui-lin YANG
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Anti-Idiotypic; biosynthesis; blood; Antibodies, Viral; biosynthesis; immunology; Blotting, Western; Capsid Proteins; genetics; immunology; metabolism; Clinical Laboratory Techniques; Cloning, Molecular; methods; Electrophoresis, Polyacrylamide Gel; Enterovirus; chemistry; immunology; isolation & purification; Enzyme-Linked Immunosorbent Assay; methods; Gene Expression; Hand, Foot and Mouth Disease; virology; Humans; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity
- From: Chinese Journal of Experimental and Clinical Virology 2008;22(6):492-494
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain a recombinant purified Enterovirus 71 VPI protein and establishment of an early, rapid and accurate serological ELISA (enzyme-linked immunosorbent assay) for detection of EV71 infection.
METHODSVP1 gene was amplified by PCR and clonel into pET-21b (+) vector, the positive recombinant plasmid were transformed into E. coli BI21(DE3), and was induced with IPTG, the recombinant protein by SDS-PAGE and Western Blot assays. Finally, the recombinant purified VP1 protein was used as a coated antigen for detection of serum anti-IgM and IgG against EV71 by ELISA.
RESULTSThe purified VP1 was obtained, and it can be recognized by sera of patients with EV71 infection associated with hand-foot-mouth disease. The A values of anti-EV71 IgM and IgG were significantly elevated as compared to healthy objects and HFMD patients without EV71 infection (P < 0.05). The sensitivity and specificity of IgM to EV71 were 73% and 77% compared with the RT-PCR results, respectively;and those of IgG being 82% and 83%, respectively.
CONCLUSIONSThe recombinant protein VP1 was produced and purified, and it was proved to have a good antigenicity and could be used to develop a serological diagnosis kit for EV71 infection in the future.
