Introduction: Use of pulse oximetry devices (PODs) by healthcare workers and the general population is becoming popular, especially after coronavirus disease 2019 (COVID-19) pandemic. Use of PODs has several limitations that may lead to inaccurate readings. Hence, we planned to do a study to assess the basic working knowledge of pulse oximetry among healthcare workers and patients. Materials and methods: This was a cross-sectional descriptive study conducted among the healthcare workers and patients requiring oxygen therapy admitted to MM Medical College & Hospital, Kumarhatti-Solan. Total duration of the study was 6 months from January to June 2023. A validated previously published semi-structured simplified questionnaire containing a total of 20 questions was used to assess the knowledge of pulse oximetry among the study population. The collected data were entered into Statistical Package for the Social Sciences (SPSS) version 20 for analysis. Results: Out of 290 participants in this study, 89 (30.7%) were nurses, 76 (26.2%) were doctors, 73 were (25.2%) medical students, and 52 (17.9%) were patients. Majority, 189 (68.3%), of the participants had poor knowledge of pulse oximetry. Lack of formal training of pulse oximetry was associated with poor knowledge of pulse oximetry among healthcare workers (p-value < 0.001). Lower level of education status and socioeconomic status were negatively correlated with patients’ basic working knowledge of pulse oximetry. Conclusion: Our study noted poor basic working knowledge of pulse oximetry among healthcare workers and patients requiring home oxygen therapy. There is an utmost need of pulse oximetry training for healthcare professionals and patients to ensure quality care for patients.