DAO 117:21-29 (2015)  -  DOI: https://doi.org/10.3354/dao02921

Rapid immunochromatographic test strip to detect swimming crab Portunus trituberculatus reovirus

LiPing Zhang1,*, DengFeng Li1,*,**, LianGuo Liu1, Ge Zhang2

1School of Marine Sciences, Ningbo University, Fenghua Road, Ningbo, Zhejiang 315211, PR China
2College of Biological Science and Technology, Hunan Agriculture University, Nongda Road, Changsha, Hunan 410128, PR China
*These authors contributed equally to this work
**Corresponding author:

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.


KEY WORDS: Portunus trituberculatus · Reovirus · Rapid detection · Immunochromatographic strip · Gold nanoparticle label · Competitive assay · Egg yolk antibody (IgY)


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Cite this article as: Zhang LP, Li DF, Liu LG, Zhang G (2015) Rapid immunochromatographic test strip to detect swimming crab Portunus trituberculatus reovirus. Dis Aquat Org 117:21-29. https://doi.org/10.3354/dao02921

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