Introduction: Tuberculosis (TB) remains one of the leading infectious causes of death worldwide, and India is among the countries with the highest TB burden. TB control is facing several roadblocks in our country with the rapid development of multidrug-resistant (MDR) as well as extensively drug-resistant TB (XDR) and as an after-effect of the global COVID-19 pandemic. With the target of TB elimination by 2025 (National Tuberculosis Elimination Program, NTEP), there is a need that treating physicians in our country be well aware of MDR-TB and be able to diagnose and treat it at an appropriate time. The present study is conducted to explore the knowledge levels, attitudes, and practices concerning MDR-TB amongst healthcare professionals working in different healthcare sectors.Methods: A total of 250 allopathic medical practitioners (Bachelor of Medicine and Bachelor of Surgery [MBBS], specialists, and superspecialists) working in any sector (private or government), who are directly involved in managing any form of TB patient and are willing to undertake the assessment, were included in this online questionnaire-based survey that was circulated using various social media platforms like WhatsApp, Facebook, Linked In, and Gmail. Responses to the questionnaires created in Google Forms were analyzed by capturing data in a Microsoft Excel® spreadsheet for further statistical analysis. The data were analyzed using multiple measures of dispersion and cross-tabulations.Results: Among the 250 participants, most of the participants had encountered MDR-TB in their clinical practice, and the majority believe that MDR-TB is a rising problem. Although 88% of the participants did a GeneXpert assay before the start of anti-tubercular therapy (ATT), three-fourths of the participants knew that the assay detects the MTB genome and rifampicin resistance. MDR-TB was suspected in participants after no clinical improvement was observed after 3-6 weeks of a trial of ATT. Two-thirds of the participants knew that linezolid is currently being used as a second-line drug for the treatment of MDR-TB. The respondents in our survey mostly do not themselves treat MDR-TB and refer the patients to an MDR-TB center or a pulmonary medicine specialist. Conclusion: Healthcare practitioners (HCPs) with good knowledge levels can diagnose and treat TB patients appropriately, thus decreasing the rising MDR-TB problem, and they can educate patients and the general population about TB and the emerging MDR-TB situation. With the current level of knowledge about MDR-TB management, there is certainly an urgent need for educational and persuasive measures for the training of doctors in both the public and private sectors so as to achieve TB elimination by 2025.