Staphylococcus aureus is one of the most common pathogens that cause a wide
range of infections ranging from skin and soft tissue infections to invasive, life
threatening infections. The emergence of methicillin-resistant Staphylococcus aureus
(MRSA) substantially increased healthcare burdens associated with Staphylococcal
infections because of high morbidity and mortality and increasing the need for efficient
and cost-effective screening methods, for high-risk patients. The objective of this study
is to develop two molecular methods, real-time PCR and loop-mediated isothermal
amplification (LAMP), and validate them following Clinical Laboratory Improvement
Amendments (CLIA) and College of American Pathologists (CAP) standards. The real
time PCR assay was developed targeting mecA, mecC, nuc, and coa to detect S. aureus
and methicillin-resistance. The assay had high precision, a linear range of 104-108
CFU/ml, and 95% accuracy. The assay detects MRSA, MSSA, MR-CoNS, and MS
CoNS. The LAMP assay was developed targeting the same genes; however, its lower
limit of detection was 106 CFU/ml, which was much higher than that of the real-time
PCR assay. Additional studies are required to optimize the performance characteristics
of the LAMP assay further. Nevertheless, the real-time PCR assay developed in this
study will be useful for the detection of MRSA in a cost-effective manner
