The article describes the development and application of PCR primers to detect various erythromycin-resistant determinants in clinical isolates. The authors designed specific primers to amplify and distinguish between different mechanisms of resistance, including Erm methylases, efflux pumps, and inactivating enzymes. The PCR methodology allows for the direct sequencing of amplified products, facilitating the comparison of resistance determinants in clinical strains. The primers were tested on a variety of reference strains and clinical isolates, successfully identifying the presence of erm, msr, ere, mph, and mef determinants. The multiplex PCR approach not only detects resistance but also helps in distinguishing the nature of the resistance determinant, which is crucial for effective antibiotic therapy and resistance surveillance. The study highlights the importance of this methodology in clinical settings to guide appropriate antibiotic usage and prevent the selection of resistant strains.The article describes the development and application of PCR primers to detect various erythromycin-resistant determinants in clinical isolates. The authors designed specific primers to amplify and distinguish between different mechanisms of resistance, including Erm methylases, efflux pumps, and inactivating enzymes. The PCR methodology allows for the direct sequencing of amplified products, facilitating the comparison of resistance determinants in clinical strains. The primers were tested on a variety of reference strains and clinical isolates, successfully identifying the presence of erm, msr, ere, mph, and mef determinants. The multiplex PCR approach not only detects resistance but also helps in distinguishing the nature of the resistance determinant, which is crucial for effective antibiotic therapy and resistance surveillance. The study highlights the importance of this methodology in clinical settings to guide appropriate antibiotic usage and prevent the selection of resistant strains.