Development of an immuno-based colorimetric assay for white spot syndrome virus

作者：Loyprasert Thananimit Suchera^*; Saleedang Akrapon; Deachamag Panchalika; Waiyapoka Thanya****; Neulplub Maitee; Chotigeat Wilaiwan

来源：Biotechnology and Applied Biochemistry, 2014, 61(3): 357-362.

DOI：10.1002/bab.1169

• white spot syndrome virus (WSSV)
• immunoassay
• streptavidin magnesphere paramagnetic particles (SMPPs)
• shrimp
• binding protein
• colorimetric assay

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 x 10(4) and 1.6 x 10(7) copies mu L-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.

• 出版日期2014-6