Background: MicroRNAs are approximately 22 nucleotides (nt) in length, small non-coding RNAs (ncRNAs), playing a vital role during post-transcriptional regulation of mycobacterial infection. Few sputum microRNAs (sputum miR) were evaluated in literature for their role as potential biomarkers in diagnosing Pulmonary Tuberculosis (PTB), but the correlation with a full course of Tuberculosis (TB) treatment is yet to be ascertained. Methods: Sputum samples of forty-six PTB patients were collected for Acid Fast Bacilli (AFB) staining and sputum microRNA-144 (sputum miR144) copy analysis, before, during, and at the end of treatment as per Indian National guidelines. Twelve patients of Asthma or Chronic Obstructive Pulmonary Disease (COPD), not in infective exacerbation, were recruited as controls. Sputum miR-144 copy levels were measured by quantitative Real-Time PCR (qRT-PCR) method. pResults: The difference between the baseline sputum miR-144 copy levels of patients and that of controls was found to be significantly higher (p <0.001). Significant up-regulation of sputum miR-144 copy levels at the baseline, as compared to controls, and significant downregulation during and after the full course of treatment was noted (all p <0.001). Conclusion: The potential role of sputum miR-144 as a simple and non-invasive surrogate biomarker in the diagnosis and treatment of PTB gains importances.