Optimizing Molecular Tools for Bioaerosol Monitoring: A Case Study of Staphylococcus aureus in a Crowded Workplace

Staphylococcus aureus is a common opportunistic pathogen found in various environments, with the potential for rapid spread, especially in densely populated indoor settings. Integrating traditional microbiological monitoring with molecular techniques is critical for the timely detection and control of such pathogens. The aim of this study was (1) to monitor the presence and spread of S. aureus in a crowded occupational environment and (2) to optimize a PCR protocol with sequence specific primers (PCR-SSP) for precise identification and early detection of this microorganism and its antibiotic resistance genes. Sampling was conducted in two different places: a call center and a healthcare facility room. All samples were collected from indoor areas at two different time points (T0 and T1) in May 2025 (mean temperature: 22.5 °C; humidity: 59.5%). Microbiological techniques and molecular analysis using PCR-SSP were employed to confirm the presence of S. aureus and detect antibiotic resistance genes such as mecA. A total CFU (colony-forming unit) count of 587 was recorded at the dental clinic corridor, and a total CFU count of 2008 was recorded at the call center corridor. PCR-SSP successfully confirmed the identity of S. aureus with an amplicon size 267 bp and enabled the detection of antibiotic resistance markers, validating its use as a complementary method to traditional microbiological techniques. This study highlights the importance of combining environmental monitoring with molecular biology tools to enhance the early detection and accurate identification of microbial pathogens such as S. aureus and provide an insight for our future direction of producing biosensors for digital air monitoring in crowded workplaces.