This study aimed to perform a comparative validation of the efficiency of quantitative SYBR Green qPCR and simplex PCR identification of Salmonella spp. DNA. For this, the samples of DNA from the Salmonella typhimurium were diluted up to 10-5 in duplicate. The results showed that the same primers were effective for both simplex PCR and qPCR. It was possible to detect the bovine species up to a dilution of 10-1 using simplex PCR. For all dilutions, it was possible to obtain qPCR amplification with a minimum Ct value of 15.13 for the 10-1 dilution for Salmonella spp. Next, the SYBR Green qPCR amplicons were separated using agarose gel electrophoresis for confirmation of the amplified fragment size. The superiority of qPCR over multiplex PCR was validated in terms of sensitivity, even with the use of SYBR Green dye, suggesting the possible use for quality control in foodstuffs.