PMID- 38160910
OWN - NLM
STAT- MEDLINE
DCOM- 20240115
LR  - 20240520
IS  - 1872-7492 (Electronic)
IS  - 0168-1702 (Print)
IS  - 0168-1702 (Linking)
VI  - 340
DP  - 2024 Feb
TI  - Propidium monoazide PCR, a method to determine OsHV-1 undamaged capsids and to 
      estimate virus Lethal Dose 50.
PG  - 199307
LID - S0168-1702(23)00269-1 [pii]
LID - 10.1016/j.virusres.2023.199307 [doi]
LID - 199307
AB  - Ostreid herpes virus 1 (OsHV-1) has been classified within the 
      Malacoherpesviridae family from the Herpesvirales order. OsHV-1 is the 
      etiological agent of a contagious viral disease of Pacific oysters, C. gigas, 
      affecting also other bivalve species. Mortality rates reported associated with 
      the viral infection vary considerably between sites and countries and depend on 
      the age of affected stocks. A variant called muVar has been reported since 2008 in 
      Europe and other variants in Australia and in New Zealand last decade. These 
      variants are considered as the main causative agents of mass mortality events 
      affecting C. gigas. Presently there is no established cell line that allows for 
      the detection of infectious OsHV-1. In this context, a technique of propidium 
      monoazide (PMA) PCR was developed in order to quantify "undamaged" capsids. This 
      methodology is of interest to explore the virus infectivity. Being able to 
      quantify viral particles getting an undamaged capsid (not only an amount of viral 
      DNA) in tissue homogenates prepared from infected oysters or in seawater samples 
      can assist in the definition of a Lethal Dose (LD) 50 and gain information in the 
      experiments conducted to reproduce the viral infection. The main objectives of 
      the present study were (i) the development/optimization of a PMA PCR technique 
      for OsHV-1 detection using the best quantity of PMA and verifying its 
      effectiveness through heat treatment, (ii) the definition of the percentage of 
      undamaged capsids in four different tissue homogenates prepared from infected 
      Pacific oysters and (iii) the approach of a LD50 during experimental viral 
      infection assays on the basis of a number of undamaged capsids. Although the 
      developped PMA PCR technique was unable to determine OsHV-1 infectivity in viral 
      supensions, it could greatly improve interpretation of virus positive results 
      obtained by qPCR. This technique is not intended to replace the quantification of 
      viral DNA by qPCR, but it does make it possible to give a form of biological 
      meaning to the detection of this DNA.
CI  - Copyright (c) 2023. Published by Elsevier B.V.
FAU - Renault, Tristan
AU  - Renault T
AD  - Departement Ressources Biologiques et Environnement, Ifremer, Nantes, France. 
      Electronic address: tristan.renault@ifremer.fr.
FAU - Faury, Nicole
AU  - Faury N
AD  - ASIM, Adaptation Sante des Invertebres, Ifremer, La Tremblade, France.
FAU - Morga, Benjamin
AU  - Morga B
AD  - ASIM, Adaptation Sante des Invertebres, Ifremer, La Tremblade, France.
LA  - eng
PT  - Journal Article
PT  - Review
DEP - 20240104
PL  - Netherlands
TA  - Virus Res
JT  - Virus research
JID - 8410979
RN  - 0 (DNA, Viral)
RN  - 0 (propidium monoazide)
RN  - 0 (Azides)
RN  - 36015-30-2 (Propidium)
SB  - IM
MH  - Animals
MH  - *Herpesviridae/genetics
MH  - DNA, Viral/genetics
MH  - Capsid
MH  - Lethal Dose 50
MH  - *Crassostrea/genetics
MH  - Polymerase Chain Reaction
MH  - *Virus Diseases
MH  - *Azides
MH  - Propidium/*analogs & derivatives
PMC - PMC10800765
OTO - NOTNLM
OT  - Lethal Dose 50
OT  - Mortality
OT  - OsHV-1
OT  - PCR
OT  - Pacific oyster
OT  - Propidium monoazide
OT  - Undamaged capsids
COIS- Declaration of Competing Interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2024/01/02 11:42
MHDA- 2024/01/15 12:42
PMCR- 2024/01/04
CRDT- 2023/12/31 19:16
PHST- 2023/08/07 00:00 [received]
PHST- 2023/12/27 00:00 [revised]
PHST- 2023/12/28 00:00 [accepted]
PHST- 2024/01/15 12:42 [medline]
PHST- 2024/01/02 11:42 [pubmed]
PHST- 2023/12/31 19:16 [entrez]
PHST- 2024/01/04 00:00 [pmc-release]
AID - S0168-1702(23)00269-1 [pii]
AID - 199307 [pii]
AID - 10.1016/j.virusres.2023.199307 [doi]
PST - ppublish
SO  - Virus Res. 2024 Feb;340:199307. doi: 10.1016/j.virusres.2023.199307. Epub 2024 
      Jan 4.