Detection of isothermally amplified ostreid herpesvirus 1 DNA in Pacific oyster (Crassostrea gigas) using a miniaturised electrochemical biosensor
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Furones, M. Dolors
O'Sullivan, Ciara K.
Given the threat that ostreid herpesvirus 1 (OsHV-1) poses to shellfish aquaculture, the need for rapid, user-friendly and cost-effective methods to detect this marine pathogen and minimise its impact is evident. In this work, an electrochemical biosensor for the detection of OsHV-1 based on isothermal recombinase polymerase amplification (RPA) was developed. The system was first tested and optimised on maleimide microtitre plates as a proof-of-concept, before being implemented on miniaturised gold electrodes. Amperometric detection of the isothermally amplified product was achieved through a sandwich hybridisation assay with an immobilised thiolated capture probe and a horseradish peroxidase (HRP)-labelled reporter probe. Calibration curves were constructed using PCR-amplified OsHV-1 DNA, achieving a limit of detection of 207 OsHV-1 target copies. The biosensor was applied to the analysis of 16 oyster samples from an infectivity experiment, and results were compared with those obtained by qPCR analysis, showing a strong degree of correlation (r = 0.988). The simplicity, rapidity, cost-effectiveness and potential for in-situ testing with the developed biosensor provide a valuable tool for the detection of OsHV-1 in aquaculture facilities, improving their management.
639 - Caça. Pesca. Piscicultura
Is part of
Toldrà, Anna, M. Dolors Furones, Ciara K. O'Sullivan, and Mònica Campàs. 2020. "Detection Of Isothermally Amplified Ostreid Herpesvirus 1 DNA In Pacific Oyster (Crassostrea Gigas) Using A Miniaturised Electrochemical Biosensor". Talanta 207: 120308. Elsevier BV. doi:10.1016/j.talanta.2019.120308.
Grant agreement number
EC/H2020/678589/EU/Preventing and mitigating farmed bivalve diseases/VIVALDI
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