Detection of Viable Salmonella Typhimurium and Staphylococcus aureus in Coalho Cheese by Real-Time PCR

Abstract Real-time PCR (qPCR) has been used for rapid identification of Salmonella Typhimurium and Staphylococcus aureus in dairy foods, but is unable to differentiate viable and unviable pathogens. Ethidium bromide monoazide (EMA), a DNA-intercalating agent, can detect only viable cells because selectively enter cells considered unviable and bind to their DNA, inhibiting its amplification during qPCR. The objective was to establish a protocol for detection of viable Salmonella Typhimurium and S. aureus, experimentally inoculated in coalho cheese, by the use of EMA combined with qPCR. The prot...