The aim of this study was to explore perceptions and knowledge of source isolation among hospitalised patients colonised or infected with multi-resistant organisms, to identify if information provided and delivery method are helpful and appropriate, and to identify areas for practice improvements. Purposive sampling was conducted. Between November 2019 and January 2020, bedside interviews with structured questionnaires (combining multiple-choice and free-text questions) were conducted with adult in-patients requiring isolation for multi-resistant organisms in a 180-bed metropolitan hospital in Brisbane, Australia. Data analysis included quantifying multiple-choice responses and thematic analysis of free-text responses. Thirty participants completed the interview questionnaire. Lack of awareness and understanding of multi-resistant organisms was evident. Participants reported a preference for face-to-face education (96.7%) and information brochures (86.7%), rather than phone call (33.3%) or informative video (0%). Qualitative responses revealed communication and information deficits exacerbated patients’ negative psychological impacts including embarrassment, loneliness, abandonment, confusion and fear. Participants identified that clinicians need better communication skills and knowledge of multi-resistant organisms to recognise and ameliorate the effect of source isolation on patients. In conclusion, patients in source isolation reported that they do not receive adequate information. Enhancing clinician knowledge of multi-resistant organisms and improving communication skills may help address the psychological needs of these patients.