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Abstract Background Cross-cultural competence is widely regarded to play an important role in being able to deliver appropriate and effective health care to patients with different backgrounds, race, gender orientation and cultural beliefs. This study aims to assess how medical students feel about their comfort, knowledge, and skill level in handling a diverse patient population using a validated questionnaire. Methods This study was carried out over a period of three weeks from July 5th to July 26th of 2021, in the Aga Khan University Hospital, Karachi, Pakistan. All medical students who fulfilled the eligibility criteria and gave informed consent were included in the study. A modified version of the Harvard cross-cultural care survey was used to assess the medical students’ comfort, knowledge and skill level in a variety of circumstances related to patients with different backgrounds and cultures. Descriptive statistical analysis of the questionnaire items was carried out. We reported frequencies and percentages for gender and year of study. For the questionnaire items, we reported mean, assuming that our Likert scale had equivariant intervals. Furthermore, multivariate analysis between demographics and themes was carried out. A p-value of