Although there are plenty public toilet facilities in the holy sites during Hajj, the presence of the pathogenic bacteria on the doorknobs poses a potential risk to the pilgrims. The present study aimed to identify microbial contaminants and evaluate the prevalence of MRSA and PVL strains from doorknobs surfaces of the toilets in Arafat, Muzdalifah and Mina places. Bacterial contamination was performed by swab method from 224 randomly selected toilets. Identification was done using standard microbiological methods and further confirmed using the API 20E and VITEK 2 Compact 15 identification system. The mecA and PVL genes of Staphylococcus isolates were detected by PCR. Contamination was detected in (78.3%) of doorknobs. The highest number of contamination was in Muzdalifah (100%) followed by Arafat (73.3%). The total number of Gram positive and Gram negative bacteria was (49.2%) and (35.0%) respectively. High rate of isolates identified was Staphylococcus aureus (S. aureus)(22%) followed by Coagulase negative Staphylococci (CoNS) (17.3%) and acinetobacter (10%). Out of 42 S. aureus isolates (16.7%) were found to be MRSA (positive for mecA genes) and (31 %) were positive for PVL. The study highlights the fact that bacterial contamination on doorknobs of toilets serve as a source for potential community infections. Therefore proper cleaning and effectiveness of hand washing hygiene during hajj are essential.