Abstract: Urinary tract infections (UTI) are the infection caused by microbes and women are especially prone to it than men due to anatomical reasons. Uropathogenic S.aureus have developed resistant to presently prescribed antibiotics viz., Trimethoprim and cotrimoxazole. S.aureus clinical isolates were utilized in this study.Antibacterial activity for selected phytochemicals was studied.Plasmid DNA isolation and PCR amplification of dfrAgene of DHFR S.aureus was done. In silico studies were performed to analyze the binding efficiencyof various phytochemicals and antibiotics on the DHFR protein of S.aureus.