Background: Healthcare-associated infections remain a significant threat in resource-limited settings. Standard precautions including hand hygiene, personal protective equipment usage, and safe injection practices are critical, yet often under-implemented. This study examined healthcare workers adherence to infection prevention and control (IPC) at Tiko District Hospital in Cameroon. Methods: This cross-sectional, hospital-based survey was conducted from November 2024 to June 2025. A sample of 80 workers completed the structured questionnaires covering demographics, infection control prevention adherence, influencing factors, and barriers. Data were entered into Microsoft Excel and analyzed using SPSS version 20. Descriptive statistics and chi-square tests (α = 0.05) were applied to assess adherence and its associations. Results: An overall 65% good adherence was recorded among the participants. High compliance was observed with sharps disposal and glove usage. Moderate adherence was noted in gown use and respiratory hygiene. Positive determinants of good compliance included greater years of experience (p < 0.05), regular IPC training (p < 0.01), and strong leadership support (p < 0.01). Conclusion: There exist several barriers to healthcare workers adherence to infection prevention control. While baseline notification of IPC adherence is moderate to high among nurses at Tiko District Hospital, specific precautions especially gowning and respiratory hygiene remain under-practiced. Experience, ongoing education, and leadership commitment significantly enhance compliance; resource constraints and workload impede it.