[Expression of the hepatitis Delta antigen in prokaryotic cell and evaluation of its application as an EIA diagnostic reagent].
BACKGROUND
To construct the pRSETB-HDAg recombinant expression plasmid and to obtain soluble hepatitis D virus antigen with high biological and antigenic activity.


METHODS
HDAg gene fragment was inserted into fusion expression pRSET B vector that includes T7 promoter and a polyhistidine tag. The recombinant plasmid was transformed into host bacterium BL21 after induction with IPTG. The expression supernatant was purified by chelating affinity chromatography and the recombinant HDAg antigenic activity was detected by EIA.


RESULTS
EIA detection using the recombinant HDAg showed strong positive reaction with hepatitis D patients sera. The positive rates of the EIA, compared with HDAg from USA and Hua Mei EIA kit in detecting 26 cases of anti-HDV positive reference sera, were 100%, 96.15% and 100%, respectively.


CONCLUSIONS
Recombinant plasmid for HDAg with good antigenicity was successfully constructed and could be used as hepatitis D antibody detection reagent.
Jiang
Yong-zhen
YZ
Research Laboratory of Hepatitis, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Zhang
Ming-cheng
MC
Tian
Rui-guang
RG
Lu
Jian
J
Zhang
Wen-ying
WY
Bi
Sheng-li
SL
D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D035923:Hepatitis delta Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007124:Immunoenzyme Techniques; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism
D035923:Hepatitis delta Antigens; D011994:Recombinant Proteins