β-Lactamase-Responsive Probe for Rapid and Non-invasive Detection of Tuberculosis through Induced Breath Analysis

Abstract Pulmonary tuberculosis (PTB) remains a major global public health challenge. Existing diagnostic approaches are generally limited by suboptimal sensitivity, prolonged turnaround times, high costs, and reliance on sputum samples. These constraints hinder large-scale implementation in resource-limited settings and substantially impede early screening and timely intervention. Targeting the characteristic expression of β-lactamase in Mycobacterium tuberculosis, this study employed systematic screening of a molecular compound library and identified cefapirin sodium as a specific molecular probe. Upon selective hydrolysis by β-lactamase, the probe releases hydrogen sulfide (H 2 S), which can be detected using a highly sensitive H 2 S sensing system, thereby enabling rapid and noninvasive identification of the pathogen. Gas chromatography-mass spectrometry (GC-MS) and colorimetric assays were used to validate the specificity of the enzymatic reaction and to confirm the accuracy of the generated product, collectively demonstrating the feasibility of the proposed detection platform. This method offers several advantages, including noninvasiveness, rapid response, and low cost, making it particularly suitable for application in resource-constrained regions. It provides a promising new strategy for early, point-of-care, and large-scale screening of pulmonary tuberculosis, with important implications for improving TB control efforts in high-burden settings.