Utilizing multiplex PCR as a molecular method to screen for Staphylococcus aureus pathogenicity and resistance

Rajini, Ingelela Marian and Pandey, LK (2025) Utilizing multiplex PCR as a molecular method to screen for Staphylococcus aureus pathogenicity and resistance. International Journal of Science and Research Archive, 16 (1). pp. 1023-1041. ISSN 2582-8185

Antibiotic resistance is becoming an increasingly pressing topic as Staphylococcus aureus continues to be a major human pathogen that causes a broad variety of diseases. An effective strategy for monitoring and treating S. aureus requires molecular detection and identification of resistance and virulence genes. In order to discover important virulence and antibiotic resistance genes in Staphylococcus aureus quickly and simultaneously, this research sought to improve a multiplex PCR technique. Using two sets of multiplex primers targeting 10 genes (sea, seb, sec, sed, saw, femA, eta, etb, tst, and mecA), a total of two hundred S. aureus strains were examined. Exactness and absence of nonspecific bands were achieved by meticulous optimization of the amplification settings. Restrictive fragment length polymorphism (RFLP) analysis allowed us to verify the PCR results' authenticity by digesting each gene with a particular enzyme to produce distinctive fragment sizes. Consistent findings from both multiplex PCR and individual gene screening proved that the test was reliable.