Detection of isothermally amplified ostreid herpesvirus 1 DNA in Pacific oyster (Crassostrea gigas) using a miniaturised electrochemical biosensor

• Published in: Talanta
• Main Authors: Toldrà, Anna, Furones, M. Dolors, O'Sullivan, Ciara K., Campàs, Mònica
• Other Authors: Producció Animal, Aigües Marines i Continentals, Aqüicultura
• Format: Article in Journal/Newspaper
• Language: English
• Published: Elsevier 2019
• Subjects: 639; Pacific; Crassostrea gigas; Pacific oyster
• Online Access: https://hdl.handle.net/20.500.12327/493; https://doi.org/10.1016/j.talanta.2019.120308

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. info:eu-repo/semantics/acceptedVersion