title: Development of an immuno-based colorimetric assay for white spot syndrome virus.
abstract: White spot syndrome virus (WSSV) is a major cause of infectious disease in cultured shrimp. A fast and reliable method for detecting and monitoring the amount of WSSV during farming would be extremely useful. This work describes a sandwich immunoassay that uses anti-GST-VP26, a WSSV-binding protein (WBP), and modified streptavidin magnesphere paramagnetic particles (SMPPs) to develop the technique. The WBP was immobilized on SMPPs and later bound to different copies of WSSV. The binding was detected using anti-GST-VP26 conjugated to alkaline phosphatase. This enzymatic reaction successfully changed the test solution to a concentration-dependent yellow color that was measured at 405 nm. The sensitivity of this method was between 1.6 × 10(4) and 1.6 × 10(7) copies µL(-1) of WSSV. In this study, the color for detection and semiquantitative analysis is easily observed and measured and can lead to the development of a test kit for screening WSSV during shrimp farming.
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand; Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
authors.forename: Suchera
authors.initials: S
authors.lastname: Loyprasert-Thananimit
authors.forename: Akrapon
authors.initials: A
authors.lastname: Saleedang
authors.forename: Panchalika
authors.initials: P
authors.lastname: Deachamag
authors.forename: Thanyaporn
authors.initials: T
authors.lastname: Waiyapoka
authors.forename: Maitee
authors.initials: M
authors.lastname: Neulplub
authors.forename: Wilaiwan
authors.initials: W
authors.lastname: Chotigeat
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D003124:Colorimetry / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D046848:White spot syndrome virus 1 / Q000276:immunology* / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal
keywords: binding protein; colorimetric assay; immunoassay; shrimp; streptavidin magnesphere paramagnetic particles (SMPPs); white spot syndrome virus (WSSV)