Virus-like particle-based immunoglobulin M capture enzyme-linked immunosorbent assay for the detection of IgM antibodies against Chikungunya virus.
- Author:
Jian-dong LI
;
Quan-fu ZHANG
;
Shuo ZHANG
;
Chuan LI
;
Qin-zhi LIU
;
Mi-fang LIANG
;
De-xin LI
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
blood;
Chikungunya Fever;
blood;
diagnosis;
virology;
Chikungunya virus;
immunology;
isolation & purification;
Enzyme-Linked Immunosorbent Assay;
methods;
Humans;
Immunoglobulin M;
blood;
Mice
- From:
Chinese Journal of Virology
2014;30(6):599-604
- CountryChina
- Language:Chinese
-
Abstract:
To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
