Purpose: This study explored emergency nurses’ perception of and experiences with infection control in the emergency department (ED). Methods: This qualitative study employed focus group interviews and conventional content analysis. The participants were 19 nurses working in the ED of three general hospitals in the metropolitan area, and the data collection period was from July 1, 2024, to August 30, 2024. Four focus group interviews were conducted with each group consisting of 4~5 participants. Interviews were conducted for approximately 80 minutes per group. The interviews were audio-recorded and transcribed verbatim immediately afterward. Data were analyzed using conventional content analysis. Results: A total of 167 meaningful statements were extracted from the interviews. Five main themes and 15 subthemes were identified through interview analysis. 1) Limited information with “Limited information at the entrance of the ED” and “Challenges in sharing information”; 2) The gap between guidelines and reality with “Difficulty in adhering to guidelines due to a shortage of isolation rooms”, “Underestimating protective isolation”, “Skepticism due to practical limitations” and, “The need for infection control guidelines to fit ED circumstances”; 3) Application of guidelines that deviated from the principles with “Different nursing practice based on visual judgment”, “Guidelines that were overlooked in emergency situations” and, “Justifying the avoidance of tedious work by claiming to help prevent infection”; 4) Insensitivity to infection with “Performing only pretended infection control”, “Overlooking the importance of protective equipment”, and “Being became insensitive to infection”; and 5) The need to strengthen the organizational culture of infection control with “Interpreting the same guideline differently by individuals”, “The need for support from senior nurses to comply with the guidelines”, and “The need for practical training that reflects the reality of the ED”. Conclusion: ED nurses faced challenges in complying with infection control guidelines in the ED. These challenges were attributed to a combination of physical, structural, and personal factors. A unique feature of this study is its emphasis on the organizational culture of infection control. To improve compliance with infection control guidelines among emergency nurses, it is imperative to improve their physical and structural environment and provide regular practical training. This approach aims to foster an organizational culture that places a strong emphasis on infection control.