Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water

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• 作者：Thanyaporn Waiyapoka ; Panchalika Deachamag…
• 关键词：White spot syndrome virus (WSSV) ; WSSV binding protein (WBP) ; Biosensor ; Impedance ; Label ; free immunosensor ; Shrimp cultivation
• 刊名：Applied Biochemistry and Biotechnology
• 出版年：2015
• 出版时间：October 2015
• 年：2015
• 卷：177
• 期：4
• 页码：821-830
• 全文大小：636 KB
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• 作者单位：Thanyaporn Waiyapoka (1)
  Panchalika Deachamag (2)
  Wilaiwan Chotigeat (1) (2)
  Nittaya Bunsanong (1)
  Proespichaya Kanatharana (3)
  Panote Thavarungkul (3)
  Suchera Loyprasert-Thananimit (1) (2)
  
  1. Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand
  2. Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University (PSU), Hat Yai, Songkhla, 90112, Thailand
  3. Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Biochemistry
  
• 出版者：Humana Press Inc.
• ISSN：1559-0291

文摘

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 × 101-.6 × 106 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 (80-10 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation. Keywords White spot syndrome virus (WSSV) WSSV binding protein (WBP) Biosensor Impedance Label-free immunosensor Shrimp cultivation