Introduction: Fungal secondary metabolites are a diverse group of bioactive compounds; in which many have been reported as bio-therapeutics. Objective: The present study, relates to fungal secondary metabolites produced by filamentous fungi and screened against 21 clinical isolates of Staphylococcus aureus procured from Shri Dharmasthala Manjunatheshwara Medical College and Hospital Dharwad, Karnataka. Methods: Antibiogram assay was performed for these clinical isolates against Methicillin, Chloramphenicol, Vancomycin, Oxacillin and Clindamycin by Kirby Bauer__ampersandsignrsquo;s method. The zones were measured and compared to the Clinical and Laboratory Standard__ampersandsignrsquo;s Institute chart. Results: The resistance pattern observed for Methicillin-90.47%, Chloramphenicol-4.76%, Vancomycin-14.28%, Oxacillin-80.95% and Clindamycin-9.5%. We infer that, out of 21 interactions, 5 distance, 7 overgrowth, 4 contact and 5 zone-line inhibitions were observed. The zone of inhibition range observed was 10-17 mm against all 21 S. aureus strains after 18 hr incubation at 37 __ampersandsigndeg;C on Mueller Hinton Agar plates. Bio-activity against S. aureus was noticed for F4 isolate between the 4-6th day of incubation and not on 7th day or later. The maximum inhibition zone observed for ethyl acetate:water fraction. Conclusion: The isolated compound from P. polonicum showed potent activity against the Methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates has not been used against any MRSA or Methicillin-resistant Staphylococcus aureus (VRSA) clinical isolates.