Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 173, Issue 1, Pages 75-84Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2011.01.010
Keywords
White spot syndrome virus; Affinity biosensor; Trace analysis; Shrimp pond water; Capacitance
Funding
- Thailand Research Fund
- Center for Innovation in Chemistry
- Postgraduate Education and Research Program in Chemistry (PERCH-CIC)
- National Research University Project of Thailand
- office of the Higher Education Commission
- Trace Analysis and Biosensor Research Center (TAB-RC)
- Graduate School
- Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand
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Water is one major pathways by which the white spot syndrome virus (WSSV) pathogen enters aquaculture facilities. This paper describes the production and use of a capacitive biosensor for the quantitative detection of as little as 1 copy/mu l of WSSV in shrimp pond water. A glutathione-S-transferase tag for white spot binding protein (GST-WBP) was immobilized on a gold electrode through a self-assembled monolayer. Binding between WSSV and the immobilized GST-WBP was directly detected by a capacitance measurement. Under optimum conditions, the capacitive biosensor detected WSSV over a wide linear range of between 1 and 1 x 10(5) copies/mu l. The system was highly selective for WSSV. One analysis cycle required only 20-25 min of analysis time and 25 min of regeneration time. The capacitive biosensor was applied to analyze WSSV concentration in eight shrimp pond water samples and the results were in good agreement with those obtained by a real time quantitative polymerase chain reaction (real-time PCR) method (P > 0.05). The immobilized GST-WBP provided and could be reused for up to 39 analysis cycles for one electrode preparation with a relative standard deviation (RSD) of 2.4% and a good reproducibility of residual activity (95.8 +/- 2.3%). The appealing performance of this biosensor indicated that it had great potential for an accurate very sensitive, quantitative, detection method for WSSV. (C) 2011 Elsevier B.V. All rights reserved.
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