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Staphylococcus pseudintermedius is a significant bacterial pathogen that frequently colonizes different body sites and mucous membranes of pets. The objectives of the cross-sectional study were to estimate the prevalence, antimicrobial resistance pattern, and detection of diverse resistance as well as virulence genes of S. pseudintermedius in cats. A standard bacteriological method, species-specific gene and different antimicrobial resistance as well as virulence genes were confirmed by PCR assay. A total of 233 swab samples were collected from different body sites of 102 cats, among them 146 swabs from 73 healthy cats, and 87 from 29 diseased cats. Overall, prevalence of S. pseudintermedius in cats was 12.01%, while dermatitis and otitis affected cats were 26.08% and 33.33%, respectively. The highest antimicrobial resistance was observed against penicillin (96.42%) followed by streptomycin (85.71%) and erythromycin (78.57%). Moreover, 89.28% of S. pseudintermedius isolates exhibit multi-drug resistance (MDR) (≥ 3 classes' antimicrobial resistant). In addition, 17.86% isolates harbored the mec A gene; thus, were classified as methicillin-resistant S. pseudintermedius (MRSP). Furthermore, the erythromycin resistance genes erm A and erm B were harbored by 25% and 10.71% of isolates, while 42.86% and 17.86% of isolates carried tet K and tet L (tetracycline resistance) genes, respectively. In virulence profiling, 32.14% ( sea ) and 10.71% ( seb ) of isolates were found positive for enterotoxin genes, whereas, the toxic shock syndrome toxin-1 ( tst- 1) gene and the Panton-Valentine leukocidin gene ( pvl ) were detected in 25% and 14.29% of isolates, respectively. To our knowledge, this is the first report of cats in Bangladesh for MDR S. pseudintermedius , MRSP, and their virulence profiling.