title: Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.
abstract: White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Thanyaporn
authors.initials: T
authors.lastname: Waiyapoka
authors.affiliation: Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University (PSU), Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Panchalika
authors.initials: P
authors.lastname: Deachamag
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand.|Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University (PSU), Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Wilaiwan
authors.initials: W
authors.lastname: Chotigeat
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Nittaya
authors.initials: N
authors.lastname: Bunsanong
authors.affiliation: Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Proespichaya
authors.initials: P
authors.lastname: Kanatharana
authors.affiliation: Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand.
authors.forename: Panote
authors.initials: P
authors.lastname: Thavarungkul
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand. s.thananimit@yahoo.com.|Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University (PSU), Hat Yai, Songkhla, 90112, Thailand. s.thananimit@yahoo.com.
authors.forename: Suchera
authors.initials: S
authors.lastname: Loyprasert-Thananimit
mesh_terms: D000818:Animals; D017756:Aquaculture; D015374:Biosensing Techniques / Q000379:methods*; D033364:Decapoda / Q000254:growth & development* / Q000821:virology*; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D014867:Water; D046848:White spot syndrome virus 1 / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D014867:Water
keywords: Biosensor; Impedance; Label-free immunosensor; Shrimp cultivation; WSSV binding protein (WBP); White spot syndrome virus (WSSV)