Primer Design and Optimization of Polymerase Chain Reaction Method for Detection of the mecA Gene in Methicillin-Resistant Staphylococcus aureus

Burhannuddin Burhannuddin, Ida Ayu Eka Anindita Prameswari, Ida Bagus Oka Suyasa, I Gusti Ngurah Dwija Putra

Abstract


Methicillin-Resistant Staphylococcus aureus (MRSA) is one of the main causes of nosocomial infections and exhibits resistance to beta-lactam antibiotics due to the presence of the mecA gene. Detection of MRSA is generally performed using traditional methods. However, these approaches have several limitations. With technological advancements, the Polymerase Chain Reaction (PCR) method has become a more effective option, as it can detect the presence of target DNA more quickly, accurately, and specifically. This study aimed to design specific primers and optimize the PCR method for detecting the mecA gene in MRSA. The research conducted qualitative descriptive by in silico method in primer design and in vitro method in optimization of PCR. DNA samples were obtained from MRSA ATCC 33591 isolates and extracted using the PCIA method. PCR optimization was conducted with annealing temperature variations from 47,5°C to 63,6°C. The results showed that the primer pair forward 5’-TGGCTCAGGTACTGCTATCC-‘3 and reverse 5’-TGGAACTTGTTGAGCAGAGGT-‘3 successfully amplified the mecA gene fragment of 156 bp specifically at the optimal annealing temperature of 55,5°C. It can be concluded that the primer with optimal PCR reaction produced in this study can be used as a rapid and accurate detection method for identifying the mecA gene in MRSA.

Keywords


Primer Design; Optimization; Polymerase Chain Reaction; mecA gene; Methicillin-Resistant Staphylococcus aureus

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References


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DOI: https://doi.org/10.33992/icmahs.v3i1.4567

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