Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/57495
Título: | Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes |
Autor(es): | Rocha, Rui Sousa, J. Cerqueira, Laura Vieira, Maria João Almeida, Carina Azevedo, Nuno Filipe Ribeiro Pinto Oliveira |
Palavras-chave: | Listeria monocytogenes PNA-FISH Blocker probe Enrichment procedure |
Data: | Jun-2019 |
Editora: | Elsevier |
Revista: | Food Microbiology |
Citação: | Rocha, Rui; Sousa, J.; Cerqueira, Laura; Vieira, Maria João; Almeida, Carina; Azevedo, Nuno F., Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes. Food Microbiology, 80, 1-8, 2019 |
Resumo(s): | 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 24h plus 18h. 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 (0.22CFU/25g or mL) and a high level (210CFU/25g 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 0.5CFU/25g or mL of food sample. |
Tipo: | Artigo |
Descrição: | Supplementary data to this article can be found online at https:// doi.org/10.1016/j.fm.2018.12.009. |
URI: | https://hdl.handle.net/1822/57495 |
DOI: | 10.1016/j.fm.2018.12.009 |
ISSN: | 0740-0020 |
e-ISSN: | 1095-9998 |
Versão da editora: | http://www.elsevier.com/locate/issn/07400020 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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document_49375_1.pdf | 544,88 kB | Adobe PDF | Ver/Abrir |