RT info:eu-repo/semantics/article
T1 Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages
A1 Quijada, Narciso M.
A1 Fongaro, Gislaine
A1 Barardi, Célia R. M.
A1 Hernández Pérez, Marta
A1 Rodríguez Lázaro, David
K1 enteric viruses
K1 infectivity prediction
K1 PMA-PCR
K1 clam
K1 fermented sausage
K1 Microbiology
K1 Microbiología
AB The increase of foodborne viral outbreaks highlights the need for a rapid and sensitivemethod for the prediction of viral infectivity in food samples. This study assesses the useof propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCRfor RNA or DNA viruses, respectively) in the determination of viral infectivity in complexanimal-related food matrices. Clam and Spanish fermented sausage (“chorizo”) sampleswere spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) andmengovirus (vMC0). PMA-qPCR/RT-qPCR discriminated infective virus particles, withsignificant reductions (>2.7 log10 or 99.7%). Additionally, infectious HAdV-2 and vMC0were quantified by plaque assay (in plaque forming units, PFU), and compared withthose in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistentcorrelation (R2 > 0.92) was showed between PFU and GCs along serial 10-fold dilutionsin both DNA and RNA virus and in both food matrices. This study shows the use of PMAcoupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity infood samples in comparison to more expensive and time-consuming methods and forthose viruses that are not able to grow under available cell culture techniques.
PB Frontiers Media
YR 2016
FD 2016-12
LK http://hdl.handle.net/10259/4406
UL http://hdl.handle.net/10259/4406
LA eng
NO ThisstudywasfinanciallysupportedbytheRTA2014-00024-C04-01 fromtheSpanishMinistryofEconomyandInnovationand theBrazilianCNPqProjectnumber472804/2013-8,andbyCAPES/PNPD andCAPES/PDSE.
DS Repositorio Institucional de la Universidad de Burgos
RD 23-nov-2024