Optimization fnbA gene target for potential detection Staphylococcus aureus as foodborne pathogen bacteria using Polymerase Chain Reaction method

Staphylococcus aureus can be found in a wide range of areas and emerge as one of the pathogenic bacteria that cause foodborne infection. Hence, the optimization for S. aureus detection needs to be developed to produce more accurate results. This study aims to determine the optimal annealing temperature condition and the potential of fnbA primers target gene in developing S. aureus detection using the Polymerase Chain Reaction method. Primer pairs fnbA design was first synthesized and followed by the cultural growth process of S. aureus bacteria in MSA agar, producing yellow colonies. DNA isolates with a concentration of 26 ng/uL and purity 1,8 of S. aureus ATCC 25923 were used as a PCR template. The optimization stage is carried out by varying the temperature at a range of 57-61 °C resulting in the appearance of annealing bands on agar when exposed to UV light. The results showed that the primer successfully amplified the fnbA S. aureus gene with a single band, showed the brightest color at a temperature of 60 °C, and produced an amplicon 187 bp size as targeted. Thus, the optimal conditions obtained can be used for the next stage of detection in food using Real-time PCR.