A rapid amplification/detection assay for analysis of Mycobacterium tuberculosis using an isothermal and silicon bio-photonic sensor complex.

Global tuberculosis (TB) control is hampered by cost and slow or insensitive diagnostic methods to be used for TB diagnosis in clinic. Thus, TB still remains a major global health problem. The failure to rapidly and accurately diagnose of TB has posed significant challenges with consequent secondary resistance and ongoing transmission. We developed a rapid Mycobacterium tuberculosis (MTB) amplification/detection method, called MTB isothermal solid-phase amplification/detection (MTB-ISAD), that couples isothermal solid-phase amplification and a silicon biophotonics-based detection sensor to allow the simultaneous amplification and detection of MTB in a label-free and real-time manner. We validated the clinical utility of the MTB-ISAD assay by detecting MTB nucleic acid in sputum samples from 42 patients. We showed the ability of the MTB-ISAD assay to detect MTB in 42 clinical specimens, confirming that the MTB-ISAD assay is fast (<20 min), highly sensitive, accurate (>90%, 38/42), and cost-effective because it is a label-free method and does not involve thermal cycling. The MTB-ISAD assay has improved time-efficiency, affordability, and sensitivity compared with many existing methods. Therefore, it is potentially adaptable for better diagnosis across various clinical applications.