| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 3406790 | Journal of Virological Methods | 2012 | 4 Pages |
A simple gold nanoparticle (GNP) probe based assay (GNPA) that was modified from a bio-barcode assay (BCA) technique, was developed for ultra-sensitive, rapid detection of the bluetongue virus (BTV) VP7 outer-core protein. This assay captures the VP7 target antigen using the GNP probe coated with anti-VP7 polyclonal antibodies and single-stranded signal DNA. Magnetic microparticle (MMP) probes coated with anti-VP7 monoclonal antibodies were then added to form a sandwich immuno-complex. The single-stranded signal DNA coated onto the GNP probe present in the immuno-complex could then be detected by PCR and real-time fluorescence PCR using a TaqMan probe. The assay has a purified VP7 detection limit of 10−2 fg/ml which is 8 orders of magnitude greater than that of conventional antigen capture ELISAs and 1 order of magnitude more sensitive than that of a conventional BCA. These results indicate that the GNPA is a highly sensitive method for easy detection of BTV proteins and that it can be modified as needed to measure the presence of other proteins.
► A gold nanoparticles probe based assay for ultra sensitive detection of bluetongue virus VP7. ► The single-stranded signal DNA in the immuno-complex was detected by PCR and real-time PCR. ► A detection limit of 10−2 fg/ml was measured for purified VP7 of BTV.
