PMID- 31594570
OWN - NLM
STAT- MEDLINE
DCOM- 20200219
LR  - 20200219
IS  - 1873-3573 (Electronic)
IS  - 0039-9140 (Linking)
VI  - 207
DP  - 2020 Jan 15
TI  - Detection of isothermally amplified ostreid herpesvirus 1 DNA in Pacific oyster 
      (Crassostrea gigas) using a miniaturised electrochemical biosensor.
PG  - 120308
LID - S0039-9140(19)30941-5 [pii]
LID - 10.1016/j.talanta.2019.120308 [doi]
AB  - 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.
CI  - Copyright (c) 2019 Elsevier B.V. All rights reserved.
FAU - Toldra, Anna
AU  - Toldra A
AD  - IRTA, Ctra. Poble Nou Km 5.5, 43540, Sant Carles de la Rapita, Tarragona, Spain.
FAU - Furones, M Dolors
AU  - Furones MD
AD  - IRTA, Ctra. Poble Nou Km 5.5, 43540, Sant Carles de la Rapita, Tarragona, Spain.
FAU - O'Sullivan, Ciara K
AU  - O'Sullivan CK
AD  - Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Av. Paisos 
      Catalans 26, 43007, Tarragona, Spain; Institucio Catalana de Recerca i Estudis 
      Avancats (ICREA), Pg. Lluis Companys 23, 08010, Barcelona, Spain.
FAU - Campas, Monica
AU  - Campas M
AD  - IRTA, Ctra. Poble Nou Km 5.5, 43540, Sant Carles de la Rapita, Tarragona, Spain. 
      Electronic address: monica.campas@irta.cat.
LA  - eng
PT  - Journal Article
DEP - 20190902
PL  - Netherlands
TA  - Talanta
JT  - Talanta
JID - 2984816R
RN  - 0 (DNA, Viral)
RN  - Ostreid herpesvirus 1
SB  - IM
MH  - Animals
MH  - Biosensing Techniques/economics/*methods
MH  - Calibration
MH  - Calorimetry
MH  - Cost-Benefit Analysis
MH  - Crassostrea/*virology
MH  - DNA Viruses/*genetics
MH  - DNA, Viral/*analysis/*genetics
MH  - Electrochemistry
MH  - Electrodes
MH  - Limit of Detection
MH  - *Miniaturization
MH  - Nucleic Acid Amplification Techniques
MH  - *Temperature
MH  - Time Factors
OTO - NOTNLM
OT  - Crassostrea gigas
OT  - Electrochemical biosensor
OT  - Isothermal recombinase polymerase amplification (RPA)
OT  - Ostreid herpesvirus 1 (OsHV-1)
OT  - Pacific oyster
EDAT- 2019/10/09 06:00
MHDA- 2020/02/20 06:00
CRDT- 2019/10/10 06:00
PHST- 2019/05/07 00:00 [received]
PHST- 2019/08/30 00:00 [revised]
PHST- 2019/08/31 00:00 [accepted]
PHST- 2019/10/10 06:00 [entrez]
PHST- 2019/10/09 06:00 [pubmed]
PHST- 2020/02/20 06:00 [medline]
AID - S0039-9140(19)30941-5 [pii]
AID - 10.1016/j.talanta.2019.120308 [doi]
PST - ppublish
SO  - Talanta. 2020 Jan 15;207:120308. doi: 10.1016/j.talanta.2019.120308. Epub 2019 
      Sep 2.