PMID- 32917059
OWN - NLM
STAT- MEDLINE
DCOM- 20210305
LR  - 20240329
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 21
IP  - 18
DP  - 2020 Sep 9
TI  - A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust 
      Detection of the Ostreid herpesvirus 1.
LID - 10.3390/ijms21186605 [doi]
LID - 6605
AB  - The Ostreid herpesvirus 1 species 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 of 
      Ostreid herpesvirus1 (OsHV-1) and its variants infecting Crassostrea 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.
FAU - Zaczek-Moczydlowska, Maja A
AU  - Zaczek-Moczydlowska MA
AD  - Institute for Global Food Security, School of Biological Sciences, Queen's 
      University Belfast, Belfast BT9 5DL, UK.
FAU - Mohamed-Smith, Letitia
AU  - Mohamed-Smith L
AD  - Institute for Global Food Security, School of Biological Sciences, Queen's 
      University Belfast, Belfast BT9 5DL, UK.
FAU - Toldra, Anna
AU  - Toldra A
AD  - IRTA, 43540 Sant Carles de la Rapita, Spain.
FAU - Hooper, Chantelle
AU  - Hooper C
AUID- ORCID: 0000-0002-9394-5577
AD  - Centre for Environment, Fisheries and Aquaculture Science, Weymouth DT4 8UB, UK.
FAU - Campas, Monica
AU  - Campas M
AUID- ORCID: 0000-0002-1220-7100
AD  - IRTA, 43540 Sant Carles de la Rapita, Spain.
FAU - Furones, M Dolors
AU  - Furones MD
AD  - IRTA, 43540 Sant Carles de la Rapita, Spain.
FAU - Bean, Tim P
AU  - Bean TP
AUID- ORCID: 0000-0002-2544-9918
AD  - The Roslin Institute, The Royal (Dick) School of Veterinary Studies, University 
      of Edinburgh, Midlothian EH25 9RG, UK.
FAU - Campbell, Katrina
AU  - Campbell K
AD  - Institute for Global Food Security, School of Biological Sciences, Queen's 
      University Belfast, Belfast BT9 5DL, UK.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20200909
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Naphthalenesulfonates)
RN  - 63451-35-4 (trisodium 
      3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate)
RN  - EC 3.6.4.- (DNA Helicases)
RN  - LAMP assay
RN  - Ostreid herpesvirus 1
SB  - IM
MH  - Animals
MH  - Crassostrea/virology
MH  - DNA Helicases
MH  - DNA Viruses/*isolation & purification
MH  - Molecular Diagnostic Techniques/*methods
MH  - Naphthalenesulfonates
MH  - Nucleic Acid Amplification Techniques/*methods
PMC - PMC7555478
OTO - NOTNLM
OT  - C. gigas
OT  - LAMP
OT  - OsHV-1
OT  - POCT
OT  - colorimetric
OT  - early warning detection
OT  - oyster
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses or interpretation of data; in 
      the writing of the manuscript; or in the decision to publish the results.
EDAT- 2020/09/13 06:00
MHDA- 2021/03/06 06:00
PMCR- 2020/09/01
CRDT- 2020/09/12 01:02
PHST- 2020/07/21 00:00 [received]
PHST- 2020/08/28 00:00 [revised]
PHST- 2020/09/04 00:00 [accepted]
PHST- 2020/09/12 01:02 [entrez]
PHST- 2020/09/13 06:00 [pubmed]
PHST- 2021/03/06 06:00 [medline]
PHST- 2020/09/01 00:00 [pmc-release]
AID - ijms21186605 [pii]
AID - ijms-21-06605 [pii]
AID - 10.3390/ijms21186605 [doi]
PST - epublish
SO  - Int J Mol Sci. 2020 Sep 9;21(18):6605. doi: 10.3390/ijms21186605.