Peer-reviewed veterinary case report
Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection.
- Journal:
- Archives of virology
- Year:
- 2015
- Authors:
- Bai, Xiaofei et al.
- Affiliation:
- Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences · China
- Species:
- bird
Plain-English summary
Researchers have developed a new test to detect duck Tembusu virus (DTMUV), which can cause illness in ducks. They created special antibodies that can specifically identify a part of the virus called the E protein. These antibodies were tested and found to work well in a lab setting, binding only to infected duck cells and not reacting with other viruses. The new test, called an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA), is very accurate, with a sensitivity of 98% and specificity of 94%, making it a reliable option for quickly diagnosing DTMUV in birds. Overall, this new test could help in identifying and managing DTMUV infections in duck populations.
Abstract
The E protein of flaviviruses is the primary antigen that induces protective immunity, but a monoclonal antibody (mAb) against the E protein of duck Tembusu virus (DTMUV) has never been characterized. Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B11, 3B6, and 4F9, respectively. An immunofluorescence assay indicated that the mAbs could specifically bind to duck embryo fibroblast (DEF) cells infected with DTMUV and that the E protein was distributed in the cytoplasm of the infected cells. Immunoglobulin isotyping differentiated the mAbs as IgG1 (1G2, 1B11, 4F9, 1F3, and 2A5) and IgG2b (3B6). The mAbs were used to identify three epitopes, A (2A5, 1F3, and 1G2), B (1B11 and 4F9), and C (3B6) on the E protein on the basis of a competitive binding assay. By using mAbs 1F3 and 3B6, we developed an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect E antigen from clinical samples. The AC-ELISA did not react with other known pathogens, indicating that the mAbs are specific for DTMUV. Compared to RT-PCR, the specificity and sensitivity of the AC-ELISA was 94.1 % and 98.0 %, respectively. This AC-ELISA thus represents a sensitive and rapid method for detecting DTMUV infection in birds.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/25588821/