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Diseases of Aquatic Organisms

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DAO 72:101-106 (2006)  -  doi:10.3354/dao072101

A simple and rapid immunochromatographic test strip for detection of white spot syndrome virus (WSSV) of shrimp

Weerawan Sithigorngul1, Sombat Rukpratanporn2, Nilawan Pecharaburanin3, Siwaporn Longyant1, Parin Chaivisuthangkura1, Paisarn Sithigorngul1,*

1Department of Biology, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand
2Center of Excellence for Marine Biotechnology at Chulalongkorn University, National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok 10330, Thailand
3Pacific Biotech, Petchaboon 67000, Thailand
*Corresponding author. Email:

ABSTRACT: A simple strip-test kit for white spot syndrome virus (WSSV) detection was developed using monoclonal antibody W29 (against the VP28 structural protein) conjugated with colloidal gold as the detector antibody. A rabbit anti-recombinant VP28F118 (rVP28) protein antibody in combination with a W28 monoclonal antibody was used as the capture complex at the test line (T), and goat anti-mouse IgG antibody (GAM) was used as the capture antibody at the control line (C). For evidence, the ready-to-use strip was kept in a plastic case and stored in a desiccated plastic bag. A sample volume of 100 µl gill homogenate in application buffer was applied to the sample chamber at one end of the strip and allowed to flow by chromatography through the nitrocellulose membrane to the other end. In test samples containing WSSV, the virus bound to the monoclonal antibody conjugated with colloidal gold and the resulting complex was captured by the antibodies at T to give a reddish-purple band. Any unbound monoclonal antibody conjugated with colloidal gold moved across T to be captured by the GAM and formed a band at C. In samples without WSSV or with WSSV below the limit of detection of the kit, only the band at C was seen. This method was 4 times less sensitive than dot blotting, and about 2000000 times less sensitive than 1-step PCR. Nonetheless, it could be used to screen individual shrimp or pooled shrimp samples to confirm high levels of WSSV infection or WSSV disease outbreaks. The beneficial features of this kit are that simple, convenient and quick results can be obtained without the requirement of sophisticated tools or special skills.

KEY WORDS: Colloidal gold · Monoclonal antibody · Polyclonal antibody · Penaeus monodon · White spot syndrome virus · WSSV

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