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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 Results It was found that students of year 5 considered themselves more knowledgeable, comfortable and skilled in dealing with patients of different backgrounds, religions and beliefs compared to students of year 1 and had a higher average score in all of these categories which was statistically significant. Additionally, students who believed it is extremely important to practice medicine with a diverse patient population also had the highest averages in perceived knowledge, comfort and skills in dealing with patients of different sociocultural backgrounds compared to students who believed it wasn’t important at all. Conclusion This is a first of its kind study in a private medical university in Pakistan and highlights the students’ self-assessment of their competence when caring for patients from different backgrounds. This study can be used as a reference study in the region to carry out further studies and to assess and improve the gaps in medical training being provided.