2026-04-28T11:35:50Zhttps://riubu.ubu.es/oai/requestoai:riubu.ubu.es:10259/44062022-04-29T12:02:47Zcom_10259_4862com_10259_5086com_10259_2604col_10259_4863
2017-04-06T07:23:11Z
urn:hdl:10259/4406
Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages
Quijada, Narciso M.
Fongaro, Gislaine
Barardi, Célia R. M.
Hernández Pérez, Marta
Rodríguez Lázaro, David
enteric viruses
infectivity prediction
PMA-PCR
clam
fermented sausage
The increase of foodborne viral outbreaks highlights the need for a rapid and sensitive
method for the prediction of viral infectivity in food samples. This study assesses the use
of propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCR
for RNA or DNA viruses, respectively) in the determination of viral infectivity in complex
animal-related food matrices. Clam and Spanish fermented sausage (“chorizo”) samples
were spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) and
mengovirus (vMC0). PMA-qPCR/RT-qPCR discriminated infective virus particles, with
significant reductions (>2.7 log10 or 99.7%). Additionally, infectious HAdV-2 and vMC0
were quantified by plaque assay (in plaque forming units, PFU), and compared with
those in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistent
correlation (R2 > 0.92) was showed between PFU and GCs along serial 10-fold dilutions
in both DNA and RNA virus and in both food matrices. This study shows the use of PMA
coupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity in
food samples in comparison to more expensive and time-consuming methods and for
those viruses that are not able to grow under available cell culture techniques.
2017-04-06T07:23:11Z
2017-04-06T07:23:11Z
2016-12
info:eu-repo/semantics/article
http://hdl.handle.net/10259/4406
eng
Frontiers in Microbiology. 2016, V. 7, art. 2008
https://doi.org/10.3389/fmicb.2016.02008
http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
Attribution 4.0 International
Frontiers Media