Background: Methicillin-Resistant Staphylococcus aureus (MRSA) has emerged as an important pathogen of public health importance causing significant morbidity. This study aimed at evaluation of the sensitivity, specificity and efficacy of loop-mediated isothermal amplification (LAMP) assay in detection of methicillin-resistance staphylococcus aureus (MRSA) in healthcare providers at Benha University Hospitals in comparison to conventional microbiological methods. Methods: A cross-sectional study was conducted on 50 healthcare workers in the ICU in Benha University Hospital. The collected nasal swabs were cultured on mannitol salt and blood agars, the obtained colonies were gram stained. Staphylococcal colonies were conventionally identified by catalase and coagulase tests and anti-microbial sensitivity was performed to identify the S. aureus methicillin sensitivity. Simultaneously, the obtained colonies undergone DNA extraction and amplification using LAMP assay. Results: LAMP assay showed 100% sensitivity, 83.3% specificity, 86.7% positive predictive value (PPV), and 100% negative predictive value (NPV) in detecting MRSA colonies in studied subjects compared to conventional microbiological method taking the former shorter time and simpler procedure in consider. Conclusion: the use of LAMP assay along with conventional microbiological methods is considered as acceptable diagnostic strategy especially in resource-limited areas.