Introduction: Infants are at risk of Staphylococcus aureus (S. aureus) colonization and infection. The aim of this study was to investigate S. aureus and methicillin-resistant S. aureus (MRSA) colonization among infants, including the prevalence, predictors of colonization, and antibiogram. Methodology: The study was cross-sectional, and involved infants aged less than one year recruited at the Princess Marie Louise Children’s Hospital in Accra, Ghana. Sociodemographic and clinical data of the participants were gathered with a structured questionnaire. Nasal swabs were also obtained from them and bacteriologically cultured. S. aureus was confirmed with the coagulase test, and MRSA was confirmed by polymerase chain reaction (PCR) of the mecA gene. Antimicrobial susceptibility testing of S. aureus was done using the Kirby-Bauer method. Results: The carriage prevalence of S. aureus and MRSA were 34.9% (45/129) and 17.10% (22/129), respectively. Colonization with coagulase-negative Staphylococci (CoNS) was protective of both S. aureus (OR = 0.008; p < 0.001) and MRSA (OR = 0.052; p = 0.005) carriage. Maintenance of good hand hygiene prevented S. aureus carriage (OR = 0.16; p < 0.001). S. aureus resistance to antibiotics decreased across penicillin (96%), trimethoprim-sulfamethoxazole (61%), tetracycline (61%), erythromycin (39%), gentamicin (39%), fusidic acid (26%), rifampicin (17%), clindamycin (7%), and linezolid (0%); 68.8% S. aureus were multidrug resistant. Conclusions: S. aureus and MRSA prevalence were high among the infants. Colonization with CoNS and good hand hygiene maintenance were predictive of MRSA and methicillin-sensitive S. aureus (MSSA) colonization, respectively.