کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6133548 | 1593470 | 2014 | 6 صفحه PDF | دانلود رایگان |
- Provide a new approach for the quantitative detection of rabies virus nucleoprotein.
- Validation of methodology was carried out.
- Application is demonstrated in practical samples, with satisfactory results.
- The sensitivity, effective linear range, and excellent repeatability were substantially better than conventional ELISA method.
- This precise and rapid TRFIA is a suitable tool for the quality control in the process of rabies vaccine production.
Sensitive, precise and rapid detection tests are needed in the quality control of rabies vaccine for rabies virus nucleoprotein. Previous studies for quantitation of rabies virus nucleoprotein focused on enzyme-linked immunosorbent assay (ELISA). A novel immunoassay for rapid determination of rabies virus nucleoprotein in rabies vaccine was first established by time-resolved fluoroimmunoassay (TRFIA). Based on a sandwich-type immunoassay format, analytes in samples were captured by one monoclonal antibody coating in the wells and “sandwiched” by another monoclonal antibody labeled with europium chelates. The immunocomplex was retained after washing, and then adopted treatment with enhancement solution; fluorescence was then measured according to the number of europiumions dissociated. Levels of the rabies virus nucleoprotein were measured in a linear range (5-2500Â mEU/mL) with a lower limit of quantitation (0.95Â mEU/mL) under optimal conditions. The repeatability, recovery, and linearity of the immunoassay were demonstrated to be acceptable. The correlation coefficient of nucleoprotein values obtained by novel TRFIA method and ELISA method was 0.981. These results showed good correlation and confirmed that this sensitive, precise and rapid TRFIA was feasible and could be more suitable for the quality control in the process of rabies vaccine production than ELISA.
Journal: Journal of Virological Methods - Volume 206, 15 September 2014, Pages 89-94