Application of an array-ELISA for evaluation of diagnostic antigens of HFRS virus
10.3760/cma.j.issn.0254-5101.2013.08.008
- VernacularTitle:Array-ELISA 技术对肾综合征出血热病毒诊断抗原的评价
- Author:
Xiaosong ZHANG
;
Xiaoping KANG
;
Li FAN
;
Yuchang LI
;
Xiaoyan WU
;
Yu ZHANG
;
Yang ZHENG
;
Yinhui YANG
- Publication Type:Journal Article
- Keywords:
Array-ELISA;
HFRS virus;
Nucleoprotein;
Prokaryotic expression;
Specificity
- From:
Chinese Journal of Microbiology and Immunology
2013;(8):595-599
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .
