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Author(s): Rui Rocha
José M. Sousa
Laura Cerqueira
Nuno F. Azevedo
Title: Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
Issue Date: 2019
Abstract: Listeria monocytogenes is one of the most important foodborne pathogens due to the high hospitalization and mortality rates associated to an outbreak. Several new molecular methods that accelerate the identification of L. monocytogenes have been developed, however conventional culture-based methods still remain the gold standard. In this work we developed a novel Peptide Nucleic Acid Fluorescence in situ Hybridization (PNA-FISH) method for the specific detection of L. monocytogenes. The method was based on an already existing PNA probe, LmPNA1253, coupled with a novel blocker probe in a 1:2 ratio. The method was optimized for the detection of L. monocytogenes in food samples through an evaluation of several rich and selective enrichment broths. The best outcome was achieved using One Broth Listeria in a two-step enrichment of 24 h plus 18 h. For validation in food samples, ground beef, ground pork, milk, lettuce and cooked shrimp were artificially contaminated with two ranges of inoculum: a low level (0.2-2 CFU/25 g or mL) and a high level (2-10 CFU/25 g or mL). The PNA-FISH method performed well in all types of food matrices, presenting an overall accuracy of 99% and a detection limit of 0.5 CFU/25 g or mL of food sample.
Related Information: info:eu-repo/grantAgreement/Comissão de Coordenação e Desenvolvimento Regional do Norte/P2020|Norte2020-Projetos Integrados ICDT/NORTE-01-0145-FEDER-000005/LEPABE-2-ECO-INNOVATION/LEPABE-2-ECO-INNOVATION
info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/Projetos Estratégicos/UID/EQU/00511/2013 - POCI-01-0145-FEDER-006939/Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia/LEPABE
info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/P2020|COMPETE - Projetos em Todos os Domínios Científicos/PTDC/DTP-PIC/4562/2014 - POCI-01-0145-FEDER-016678/Desenvolvimento de um novo método de RING FISH codificado por cores: uma ferramenta inovadora para a deteção de patogénios resistentes a antibióticos/Coded-FISH
Document Type: Artigo em Revista Científica Internacional
Rights: restrictedAccess
Appears in Collections:FEUP - Artigo em Revista Científica Internacional

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