Evaluation of viable Mycobacterium avium subsp. paratuberculosis in milk using peptide-mediated separation and Propidium Monoazide qPCR
| Autor: | Norma Arrigoni, G. Cammi, R. Savi, Giulia Cerutti, Maria Beatrice Boniotti, Matteo Ricchi, Caterina De Cicco, Vladimir Babak, C. Garbarino, Petr Kralik |
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| Rok vydání: | 2014 |
| Předmět: |
DNA
Bacterial Azides Pasteurization Paratuberculosis Peptide Real-Time Polymerase Chain Reaction Microbiology law.invention Multinomial logistic regression model law Propidium monoazide Limit of Detection Genetics medicine Animals Humans Molecular Biology chemistry.chemical_classification Microbial Viability biology business.industry food and beverages biology.organism_classification medicine.disease Food safety Intercalating Agents Mycobacterium avium subsp. paratuberculosis Milk chemistry Genes Bacterial Food Microbiology business Oligopeptides Mycobacterium Propidium |
| Zdroj: | FEMS microbiology letters. 356(1) |
| ISSN: | 1574-6968 |
| Popis: | The causative agent of paratuberculosis in ruminants, Mycobacterium avium subsp. paratuberculosis (MAP), although still a matter of debate, has been linked with Crohn's and other human diseases. The availability of rapid methods for assessing the viability of MAP cells in food, in particular milk, could be of great use for risk management in food safety. MAP viability is generally assessed using culture techniques that require prolonged incubation periods for the growth of MAP. To differentiate between viable and nonviable MAP cells in milk samples, this study explores the combination of two already described techniques: peptide magnetic bead separation followed by Propidium Monoazide qPCR. Using an Ordinal Multinomial Logistic Regression model to analyze the results obtained after spiking milk samples with mixtures containing different percentages of viable/dead cells, we were able to assess the probability of the viability status of MAP found in milk. This model was applied to contaminated pasteurized milk to ascertain the efficacy of heat treatment in MAP killing. The method reported herein can potentially be used for direct detection of MAP viability in milk. |
| Databáze: | OpenAIRE |
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