FN Archimer Export Format
PT J
TI Development of duplex TaqMan-based real-time PCR assay for the simultaneous detection of Perkinsus olseni and P. chesapeaki in host Manila clam tissue samples
BT 
AF Itoïz, Sarah
   Perennou, Morgan
   Mouronvalle, Clara
   Derelle, Evelyne
   Le Goïc, Nelly
   Bidault, Adeline
   de Montaudouin, Xavier
   Arzul, Isabelle
   Soudant, Philippe
   Chambouvet, Aurélie
AS 1:1;2:1;3:1,2;4:1;5:1;6:1,5;7:3;8:4;9:1;10:1;
FF 1:;2:;3:;4:;5:;6:;7:;8:PDG-RBE-SGMM-LGPMM;9:;10:;
C1 Univ Brest, CNRS, IRD, Ifremer, LEMAR, F-29280 Plouzané, France
   EPHE, PSL Research University, UPVD, CNRS, USR 3278 CRIOBE, Perpignan F-66360, France
   Univ. Bordeaux, CNRS, EPOC, EPHE, UMR 5805, Station Marine, F-33120 Arcachon, France
   IFREMER, Laboratory of Genetics and Pathology, Av de Mus de Loup-17390, La Tremblade, France
   Univ Brest, CNRS, IRD, Ifremer, LEMAR, F-29280 Plouzané, France
C2 CNRS, FRANCE
   UNIV PERPIGNAN, FRANCE
   UNIV BORDEAUX, FRANCE
   IFREMER, FRANCE
   UBO, FRANCE
SI LA TREMBLADE
SE PDG-RBE-SGMM-LGPMM
UM LEMAR
IN WOS Ifremer UPR
   WOS Cotutelle UMR
   copubli-france
   copubli-univ-france
IF 2.795
TC 2
UR https://archimer.ifremer.fr/doc/00711/82263/97776.pdf
LA English
DT Article
BO Planula IV
DE ;Alveolata;Co-infection;Molluscs;Inhibitory effect;Ruditapes philippinarum;qPCR
AB The aetiological agent Perkinsus olseni is globally recognised as a major threat for shellfish production considering its wide geographical distribution across Asia, Europe, Australia and South America. Another species, Perkinsus chesapeaki, which has never been known to be associated with significant mortality events, was recently detected along French coasts infecting clam populations sporadically in association with P. olseni. Identifying potential cryptic infections affecting Ruditapes philippinarum is essential to develop appropriate host resource management strategies. Here, we developed a molecular method based on duplex real-time quantitative PCR for the simultaneous detection of these two parasites, P. olseni and P. chesapeaki, in the different clam tissues: gills, digestive gland, foot, mantle, adductor muscle and the rest of the soft body. We firstly checked the presence of possible PCR inhibitors in host tissue samples. The qPCR reactions were inhibited depending on the nature of the host organ. The mantle and the rest of the soft body have a high inhibitory effect from threshold of host gDNA concentration of 2 ng.µL−1, the adductor muscle and the foot have an intermediate inhibition of 5 ng.µL−1, and the gills and digestive gland do not show any inhibition of the qPCR reaction even at the highest host gDNA concentration of 20 ng.µL−1. Then, using the gills as a template, the suitability of the molecular technique was checked in comparison with the Ray’s Fluid Thioglycolate Medium methodology recommended by the World Organisation for Animal Health. The duplex qPCR method brought new insights and unveiled cryptic infections as the co-occurrence of P. olseni and P. chesapeaki from in situ tissue samples in contrast to the RFTM diagnosis. The development of this duplex qPCR method is a fundamental work to monitor in situ co-infections that will lead to optimised resource management and conservation strategies to deal with emerging diseases. 
PY 2021
PD SEP
SO Journal Of Invertebrate Pathology
SN 0022-2011
PU Elsevier BV
VL 184
UT 000696985900006
DI 10.1016/j.jip.2021.107603
ID 82263
ER

EF