FN Archimer Export Format
PT J
TI Optimization of PMAxx pretreatment to distinguish between human norovirus with intact and altered capsids in shellfish and sewage samples
BT 
AF RANDAZZO, Walter
   KHEZRI, Mohammad
   OLLIVIER, Joanna
   LE GUYADER, Soizick
   RODRIGUEZ-DIAZ, Jesus
   AZNAR, Rosa
   SANCHEZ, Gloria
AS 1:1,2;2:3;3:4;4:4;5:1,5;6:1,2;7:2;
FF 1:;2:;3:PDG-RBE-SGMM-LSEM;4:PDG-RBE-SGMM-LSEM;5:;6:;7:;
C1 Univ Valencia, Dept Microbiol & Ecol, Av Dr Moliner 50, E-46100 Valencia, Spain.
   CSIC, IATA, Dept Preservat & Food Safety Technol, Av Agustin Escardino 7, Valencia 46980, Spain.
   Tarbiat Modares Univ, Fac Marine Sci, Dept Seafood Proc, Noor, Iran.
   IFREMER, LSEM SG2M, Lab Microbiol, BP 21105, F-44311 Nantes 03, France.
   INCLIVA, Hosp Clin Univ, Inst Clin Res, Valencia, Spain.
C2 UNIV VALENCIA, SPAIN
   IATA CSIC, SPAIN
   UNIV TARBIAT MODARES, IRAN
   IFREMER, FRANCE
   INCLIVA, SPAIN
SI NANTES
SE PDG-RBE-SGMM-LSEM
IN WOS Ifremer jusqu'en 2018
   copubli-europe
   copubli-int-hors-europe
   copubli-sud
IF 4.006
TC 74
UR https://archimer.ifremer.fr/doc/00411/52208/52970.pdf
LA English
DT Article
DE ;Intercalating dyes;Viability PCR;Norovirus;Shellfish;Sewage;RT-qPCR
AB Shellfish contamination by human noroviruses (HuNoVs) is a serious health and economic problem. Recently an ISO procedure based on RT-qPCR for the quantitative detection of HuNoVs in shellfish has been issued, but these procedures cannot discriminate between inactivated and potentially infectious viruses. The aim of the present study was to optimize a pretreatment using PMAxx to better discriminate between intact and heat-treated HuNoVs in shellfish and sewage. To this end, the optimal conditions (30 min incubation with 100 μM of PMAxx and 0.5% of Triton, and double photoactivation) were applied to mussels, oysters and cockles artificially inoculated with thermally-inactivated (99 °C for 5 min) HuNoV GI and GII. This pretreatment reduced the signal of thermally-inactivated HuNoV GI in cockles and HuNoV GII in mussels by > 3 log. Additionally, this pretreatment reduced the signal of thermally-inactivated HuNoV GI and GII between 1 and 1.5 log in oysters. Thermal inactivation of HuNoV GI and GII in PBS, sewage and bioaccumulated oysters was also evaluated by the PMAxx-Triton pretreatment. Results showed significant differences between reductions observed in the control and PMAxx-treated samples in PBS following treatment at 72 and 95 °C for 15 min. In sewage, the RT-qPCR signal of HuNoV GI was completely removed by the PMAxx pretreatment after heating at 72 and 95 °C, while the RT-qPCR signal for HuNoV GII was completely eliminated only at 95 °C. Finally, the PMAxx-Triton pretreatment was applied to naturally contaminated sewage and oysters, resulting in most of the HuNoV genomes quantified in sewage and oyster samples (12 out of 17) corresponding to undamaged capsids. Although this procedure may still overestimate infectivity, the PMAxx-Triton pretreatment represents a step forward to better interpret the quantification of intact HuNoVs in complex matrices, such as sewage and shellfish, and it could certainly be included in the procedures based on RT-qPCR. 
PY 2018
PD FEB
SO International Journal Of Food Microbiology
SN 0168-1605
PU Elsevier Science Bv
VL 266
UT 000424185900001
BP 1
EP 7
DI 10.1016/j.ijfoodmicro.2017.11.011
ID 52208
ER

EF