[[Virus-like particle-based immunoglobulin M capture enzyme-linked immunosorbent assay for the detection of IgM antibodies against Chikungunya virus].

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.

• Authors: Jian-dong Li, Quan-fu Zhang, Shuo Zhang, Chuan Li, Qin-zhi Liu, Mi-fang Liang, De-xin Li
• Journal: Bing du xue bao = Chinese journal of virology (Bing Du Xue Bao) Vol. 30 Issue 6 Pg. 599-604 (Nov 2014) ISSN: 1000-8721 [Print] China
• PMID: 25868272 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Viral
• Immunoglobulin M

Topics

• Animals
• Antibodies, Viral (blood)
• Chikungunya Fever (blood, diagnosis, virology)
• Chikungunya virus (immunology, isolation & purification)
• Enzyme-Linked Immunosorbent Assay (methods)
• Humans
• Immunoglobulin M (blood)
• Mice