Aim The aim of this study was to investigate the patterns and determinants of drug resistance to tuberculosis (TB) in a large population from Makkah, Saudi Arabia. Methods A retrospective, cross-sectional cohort study was conducted on all patients with TB who were referred to the National Tuberculosis Prevention Program in Makkah, Saudi Arabia, between January 2016 and September 2020. For each TB case, demographic data were collected in addition to the results of drug susceptibility testing (DST) for anti-TB drugs. The Statistical Package for Social Sciences (SPSS) software for Windows, version 23 (IBM Corporation, Armonk, NY, USA), was used for the statistical analysis. Results A total of 472 TB-confirmed cases were included in the analysis. The mean ± standard deviation of the age was 38.5 ± 17.7 years. The vast majority of patients were male (62.7%) and had pulmonary TB (91.7%). Only a small proportion of the patients with TB had diabetes mellitus (8.5%). Overall, the prevalence of monodrug-resistant TB ranged from 2.1% to 3.4%. Specifically, the prevalence of monodrug-resistant TB to isoniazid and streptomycin was ranked first and was equal to 3.4%. Pyrazinamide had the lowest prevalence of monodrug-resistant TB (2.1%). The prevalence of polydrug-resistant TB (PDR-TB) and multidrug-resistant TB (MDR-TB) was 1.5%. In the univariate analysis, sex (male) was the only sociodemographic factor that significantly correlated with a higher prevalence of MDR-TB. Conclusions This is the second study from Makkah to analyze the prevalence and associated risk factors of MDR-TB among patients from Makkah. Our data demonstrated that the prevalence of monodrug-resistant TB and MDR-TB was low (2.1%-3.4% and 1.5%, respectively). Diabetes mellitus was not a substantial factor correlated with a higher occurrence of MDR-TB. Additional epidemiologic studies are required to validate our results.