Development of a double antibody sandwich ELISA for detection of Chikungunya virus antigen
10.3760/cma.j.issn.1003-9279.2017.02.014
- VernacularTitle: 检测基孔肯雅病毒抗原方法的建立与初步评价
- Author:
Xiaoyue XING
1
;
Wei WU
;
Shuo ZHANG
;
Quanfu ZHANG
;
Chuan LI
;
Mifang LIANG
;
Jiandong LI
;
Dexin LI
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
- Publication Type:Journal Article
- Keywords:
Chikungunya virus;
Virus like particle;
Double antibody sandwich ELISA
- From:
Chinese Journal of Experimental and Clinical Virology
2017;31(2):148-152
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for detection of chikungunya virus(CHIKV) antigen.
Methods:CHIKV virus like particle(VLP), that contains all structural proteins, was prepared by baculovirus expression system. Mice and rabbits were immunized with the VLP to develop antibodies against CHIKV. A double antibody sandwich ELISA was established for detection of CHIKV antigens. The concentrations of the antibodies used and the reaction conditions were optimized. The detection limit and repeatability of the ELISA was evaluated.
Results:The sensitivity and specificity was estimated by 10 mimicking CHIKV sera, 90 health person sera, 40 other virus infected sera. It was show that the specificity of DAS-ELISA was 100%, the detection limit was 10 TCID50, the coefficients of variation (CV) within plate was <5%, the CV of different plates was <10%.
Conclusions:The double antibody sandwich ELISA established in this study can be used to detect the CHIKV antigen in acute phase serum of patient and provide a method for detection of CHIKV.
