Rapid immunochromatographic test strip to detect swimming crab Portunus trituberculatus reovirus

Abstract

Swimming crab reovirus (SCRV) is the causative agent of a serious disease with high mortality in cultured Portunus trituberculatus. A rapid immunochromatographic assay (ICA) was developed in a competitive assay format and optimized for the detection of SCRV. The gold probe-based ICA test comprised SCRV antigen and goat anti-chicken egg yolk antibody (IgY) sprayed onto a nitrocellulose membrane as the test line and control line, respectively. IgY-gold complexes were deposited onto the conjugate pad as detector reagents. The method showed high specificity with no cross-reactivity with other related aquatic pathogens. The detection limit of the ICA strip was 50 µg ml⁻¹. To evaluate the performance of the ICA test, the strip and an enzyme-linked immunosorbent assay (ELISA) were applied to the same samples (n = 90 crabs). The strip successfully detected SCRV in all of the artificially infected samples. Furthermore, the ICA strip and ELISA tests had high consistency (98.28%). The strip assay requires no instruments and has a detection time of less than 10 min. It is portable and easy to perform in the field. These results indicated that the developed strip could be a promising on-site tool for screening pooled crabs to confirm SCRV infection or disease outbreaks.