Abstract
Background: Tuberculosis (TB) remains a significant global health challenge, necessitating rapid and reliable diagnostic solutions. Current methods such as microscopy, culture, and PCR have limitations in resource-limited settings due to high costs and infrastructure requirements. Methods: In this study, we developed the MTB/IC LAMP-MS assay, a novel molecular diagnostic platform that integrates loop-mediated isothermal amplification (LAMP) technology with microscope (MS) detection. The assay enables TB diagnosis within 40 min, from nucleic acid extraction to result, requiring minimal equipment and simple operation for rapid and accurate detection. Results: Compared to the commercial AdvanSure TB/NTM real-time PCR kit, the MTB/IC LAMP-MS assay had a lower analytical sensitivity, with a limit of detection of 10(3) CFU/mL versus 10(2) CFU/mL for the commercial PCR kit, but showed a sensitivity of 93.3% and specificity of 100% in clinical tests. Additionally, the assay demonstrated minimal cross-reactivity with other respiratory pathogens, underscoring its robust specificity. In addition, the preloaded microchips demonstrated stable performance for up to 12 weeks at room temperature, supporting their field applicability. Conclusions: With its lower cost, simplified operation, and minimal infrastructure requirements, the MTB/IC LAMP-MS assay provides a practical and efficient solution for TB diagnosis in resource-limited settings.