Evaluation of a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk

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Published: Aug 1, 2018
Keywords:
Salmonella spp., Staphylococcus aureus, duplex PCR, raw milk

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Yuneilys Aleli Martínez-García
Centro Nacional de Sanidad Agropecuaria
Odalys Uffo-Reinoso
Centro Nacional de Sanidad Agropecuaria
Yamilka Riverón-Alemán
Centro Nacional de Sanidad Agropecuaria
José Antonio Agüero-Fernández
Centro Nacional de Sanidad Agropecuaria
Ailin Martínez-Vasallo
Centro Nacional de Sanidad Agropecuaria

Abstract

Infectious and toxigenic pathogens transmitted through food have been recognized as the cause of Foodborne Diseases (FBD). The main pathogens of concern are Salmonella spp. and Staphylococcus aureus. These bacteria are associated with acute gastrointestinal outbreaks caused by contaminated food. One of the causes of this contamination is the poor hygiene during the processing and handling of food. Foodborne outbreaks caused by milk and dairy products contaminated reach up to 6 %. The objective of the present work was to develop and standardize a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk. The principle of the method is the simultaneous amplification of the genes yfiR for Salmonella spp. and ARNr 23S for Staphylococcus aureus. The thermodynamic properties of each primer were determined and the main critical parameters of the dPCR assay were optimized. The duplex PCR demonstrated its specificity for the target microorganisms and the detection limit of the assay was up to 1 pg/μL of DNA and 102 cfu/mL in unpasteurized milk. In the milk samples analyzed, 46.7 % were contaminated with S. aureus and none of the samples were positive to Salmonella spp. The results suggest that the developed duplex PCR is sensible and specific for the simultaneous detection of both pathogens from raw milk.

Article Details

How to Cite
1.
Martínez-García YA, Uffo-Reinoso O, Riverón-Alemán Y, Agüero-Fernández JA, Martínez-Vasallo A. Evaluation of a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk. Rev. Salud Anim. [Internet]. 2018 Aug. 1 [cited 2024 Nov. 22];40(2). Available from: https://revistas.censa.edu.cu/index.php/RSA/article/view/961
Issue
Vol. 40 No. 2 (2018): mayo-agosto
Section
ARTÍCULOS ORIGINALES
National Center for Animal and Plant Health (CENSA)

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