Standardization of a rapid quadruplex PCR method for the simultaneous detection of bovine, buffalo, Salmonella spp., and Listeria monocytogenes DNA in milk
| Autor: | Mylla Christy da Silva Dufossé, J.B. Silva, Carina Martins de Moraes, Paula Fernanda Morais de Sousa, Joelson Sousa Lima, Adrianne Maria Brito Pinheiro da Rosa, Ana Paula Presley Oliveira Sampaio, Talita Bandeira Roos, Gabrielle Virgínia Ferreira Cardoso |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Salmonella
General Veterinary biology Microorganism biology.organism_classification medicine.disease_cause SF1-1100 DNA sequencing law.invention Microbiology Animal culture food pathogens chemistry.chemical_compound food safety chemistry Listeria monocytogenes law medicine Food microbiology fraud Polymerase chain reaction Bacteria DNA molecular techniques |
| Zdroj: | Arquivo Brasileiro de Medicina Veterinária e Zootecnia, Vol 73, Iss 4, Pp 781-790 (2021) Arquivo Brasileiro de Medicina Veterinária e Zootecnia v.73 n.4 2021 Arquivo brasileiro de medicina veterinária e zootecnia Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
| ISSN: | 1678-4162 |
| Popis: | The objective of the present study was to Standardize a Polymerase Chain Reaction (PCR) protocol for the authentication of bovine and buffalo milk, and to detect the presence of Salmonella spp. and Listeria monocytogenes. For this, the target DNA was extracted, mixed, and subjected to a PCR assay. Milk samples were defrauded and experimentally contaminated with microorganisms to assess the detection of target DNA at different times of cultivation, bacterial titers, and concentration of genetic material. In addition, the protocol was tested with DNA extracted directly from food, without a pre-enrichment step. The proposed quadruplex PCR showed good accuracy in identifying target DNA sequences. It was possible to simultaneously identify all DNA sequences at the time of inoculation (0h), when the samples were contaminated with 2 CFU/250mL and with 6h of culture when the initial inoculum was 1 CFU/250mL. It was also possible to directly detect DNA sequences from the food when it was inoculated with 3 CFU/mL bacteria. Thus, the proposed methodology showed satisfactory performance, optimization of the analysis time, and a potential for the detection of microorganisms at low titers, which can be used for the detection of fraud and contamination. |
| Databáze: | OpenAIRE |
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