Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 21, Issue 18, Pages -Publisher
MDPI
DOI: 10.3390/ijms21186605
Keywords
colorimetric; OsHV-1; LAMP; oyster; C; gigas; POCT; early warning detection
Funding
- European Commission [678589]
- BBSRC [BBS/E/D/20002173] Funding Source: UKRI
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TheOstreid herpesvirus 1species affects shellfish, contributing significantly to high economic losses during production. To counteract the threat related to mortality, there is a need for the development of novel point-of-care testing (POCT) that can be implemented in aquaculture production to prevent disease outbreaks. In this study, a simple, rapid and specific colorimetric loop-mediated isothermal amplification (LAMP) assay has been developed for the detection ofOstreid herpesvirus1(OsHV-1) and its variants infectingCrassostrea gigas(C. gigas). The LAMP assay has been optimized to use hydroxynaphthol blue (HNB) for visual colorimetric distinction of positive and negative templates. The effect of an additional Tte UvrD helicase enzyme used in the reaction was also evaluated with an improved reaction time of 10 min. Additionally, this study provides a robust workflow for optimization of primers for uncultured viruses using designed target plasmid when DNA availability is limited.
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