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image/svg+xml Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao Closed Access logo, derived from PLoS Open Access logo. This version with transparent background. http://commons.wikimedia.org/wiki/File:Closed_Access_logo_transparent.svg Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao Journal of Fish Dise...arrow_drop_down
image/svg+xml Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao Closed Access logo, derived from PLoS Open Access logo. This version with transparent background. http://commons.wikimedia.org/wiki/File:Closed_Access_logo_transparent.svg Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao
Journal of Fish Diseases
Article . 2021 . Peer-reviewed
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CRISPR‐based platform for rapid, sensitive and field‐deployable detection of scale drop disease virus in Asian sea bass (Lates calcarifer)

Authors: Thanwarat Sukonta; Watcharachai Meemetta; Saengchan Senapin; Saengchan Senapin; Thawatchai Chaijarasphong;

CRISPR‐based platform for rapid, sensitive and field‐deployable detection of scale drop disease virus in Asian sea bass (Lates calcarifer)

Abstract

AbstractScale drop disease virus (SDDV) is a major pathogen of Asian sea bass that has emerged in many countries across the Asia Pacific since 1992 and carries the potential to cause drastic economic losses to the aquaculture sector. The lack of an approved vaccine for SDDV necessitates timely prevention as the first line of defence against the disease, but current diagnostic platforms still face challenges that render them incompatible with field applications, particularly in resource‐limited settings. Here, we developed a novel detection platform for SDDV based on a CRISPR‐Cas12a‐based nucleic acid detection technology combined with recombinase polymerase amplification (RPA‐Cas12a). Using the viral adenosine triphosphatase (SDDV‐ATPase) gene as a target, we achieved the detection limit of 40 copies per reaction and high specificity for SDDV. The coupling with fluorescence and lateral flow readouts enables naked‐eye visualization and straightforward data interpretation requiring minimal scientific background. Compared with semi‐nested PCR in field sample evaluation, our RPA‐Cas12a assay is more sensitive and capable of detecting SDDV in asymptomatic fish. Importantly, the entire workflow can be carried out at a constant temperature of 37°C within an hour from start to finish, thus removing the need for an expensive thermal cycling apparatus and long turnaround times associated with PCR‐based methods. Therefore, owing to its high accuracy, rapidity and user‐friendliness, the developed RPA‐Cas12a platform shows the potential for diagnosis of SDDV at point of need and could be a valuable tool to help protect fish farming communities from large‐scale epidemics.

Keywords

Fish Diseases, Animals, Bass, Nucleic Acid Amplification Techniques, Sensitivity and Specificity, Iridoviridae, Perciformes

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    18
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    Top 10%
    influence
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    impulse
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    Top 10%
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
18
Top 10%
Average
Top 10%
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