Methods for diagnosis of Clostridium difficile and methods and vectors for recombinant toxin expression
US8206940B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 15, 2009 |
| Grant date | Jun 26, 2012 |
| Priority date | — |
| Expiry date | May 15, 2029 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/33
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Cell-based methods for rapid real time assay of a presence of Clostridium difficile toxin and/or cells are provided, using an assay having a toxin-enhancing antibody and a sensitive cell line carrying FcyR receptors, and kits for this assay. An ultrasensitive cell based immunocytotoxicity assay for detecting less then 1 pg/ml of C. difficile toxins in clinical samples. A TcdA-specific monoclonal antibody, AIH3, was found to significantly enhance the cytotoxicity of TcdA to macrophages and monocytes. The AIH3-dependent enhancement of glucosyltransferase activity, cytoskeleton disruption, and TNF-a production induced by TcdA was demonstrated also in RAW 264.7 cells. Methods for high level recombinant expression of C. difficile toxins in Bacillus cells, and vectors for expression, strains of Bacillus carrying the vectors are provided.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.