An enhanced immuno-dot blot assay for the detection of white spot syndrome virus in shrimp using antibody conjugated gold nanoparticles probe

White Spot Syndrome (WSS) is a worldwide ailment of penaeid shrimp, a lethal and contagious disease in shrimp caused by the White Spot Syndrome Virus (WSSV). It is essential to control; handle early findings and routine screenings under field conditions. Though PCR affirmed sensing of the causative viral nucleic acid has many advantages, for its routine use custom necessitate an advanced equipment and technical expertise. In comparison, protein-based immuno detecting methods are easier to perform by a commoner, but lack of sensitivity due to inadequate signal amplification. In this study, we focused on a sensitive immuno detection method which developed by gold nanoparticles coupled with alkaline phosphatase conjugated secondary antibody in turn recognized primary anti-serum raised against WSSV. We successfully enhanced the sensitivity of immuno-dot blot assay by 80 fold over the conventional method and visually it can be detected up to 1 ng/mL of purified WSSV. The approach described in this study is a prototype for the development of simple and inexpensive diagnostic tool will provide the routine screening of WSS in shrimp farms. (C) 2011 Elsevier B.V. All rights reserved.