Pulmonary tuberculosis, caused by Mycobacterium tuberculosis, is a significant public health issue, especially in developing countries, affecting millions of people every year. Despite the development of many antitubercular antibiotics and increased awareness of preventive methods, it is still a major cause of mortality worldwide. Vitamin D, a micronutrient known to have a major role in bone and calcium metabolism, has also shown its immunomodulatory effects to suppress mycobacterial growth. We conducted a systematic review and meta-analysis of the available evidence to explore the association between vitamin D levels and tuberculosis. We performed a systematic search for articles from inception to May 2021 in multiple databases. We included 26 studies in our qualitative synthesis and 12 studies in meta-analysis or quantitative synthesis. In our meta-analysis, we used a random-effect model to calculate the odds ratio (OR) of vitamin D deficiency in tuberculosis patients compared to the healthy controls. On pooled analysis, we found that the odds of the participants having vitamin D deficiency was 3.23 times more in tuberculosis patients compared to the healthy group (OR=3.23, CI = 1.91-5.45, p<0.0001). Thus, we concluded that there is an association between low levels of vitamin D and tuberculosis infections. We suggest conducting long-term prospective cohort studies in tuberculosis endemic countries to better understand the causal relationship between vitamin D deficiency and tuberculosis.