Background: Recent studies have shown an increasing incidence of antibiotic resistance in dacryocystitis. Management of diseases may include determining microbial agents and choosing appropriate antibiotics for treatment. Objectives: This study aimed to present the best treatments for dacryocystitis. To this end, specimens' microbiology and antibiotic susceptibility were examined in patients with dacryocystitis in the microbiology laboratory of the Kashan University of Medical Sciences. Methods: This cross-sectional study was performed on 172 patients presenting with acute and chronic dacryocystitis at the Matini Hospital, Kashan, between 2017 - 2018. Patient characteristics, culture isolates, and antimicrobial susceptibility data were collected. The PCR assay of the mecA gene was performed in all methicillin-resistant Staphylococcus isolates. Results: The most common bacteria were coagulase-negative staphylococci, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii. The majority of the isolated microbes were sensitive to rifampicin, linezolid, amikacin, and gentamicin. In Gram-negative bacilli, nine of the isolates were extended-spectrum beta-lactamase positive. The PCR test showed the frequency of mecA gene of resistant S. aureus and resistant coagulase-negative staphylococci (CoNS) isolates to be 40 and 46.3%, respectively. Conclusions: CoNS were the most frequently isolated bacteria. The highest antibiotic susceptibility was observed to rifampin, linezolid, amikacin, and gentamicin. A high percentage of CoNS carried the mecA gene.