Potential of nanopore sequencing for tuberculosis diagnosis and drug resistance detection.

Objectives: This study evaluates the effectiveness of nanopore sequencing for accurate detection of Mycobacterium tuberculosis pathogens and drug resistance mutations in clinical specimens.

Methods: A retrospective analysis of 2,421 specimens from suspected tuberculosis patients admitted to Xi'an Chest Hospital from 2022 to 2023 was conducted, with 131 specimens undergoing via real-time, fluorescence-based quantitative Polymerase Chain Reaction (qPCR), simultaneous amplification and testing RNA (RNA), Mycobacterium culture, Mycobacterium smear, and nanopore sequencing. Employing clinical tuberculosis diagnoses as the gold standard, sensitivity, specificity, positive predictive value, negative predictive value, concordance rate, and Kappa coefficient were measured for the five detection techniques. We compared nanopore sequencing with the Melting Curve method to detect drug-resistant gene mutations.

Results: Nanopore sequencing has a significantly higher sensitivity (0.786) for tuberculosis diagnosis compared to qPCR (0.411), RNA (0.411), Mycobacterium culture (0.402), and Mycobacterium smear (0.241), against the gold-standard clinical diagnosis. It also exhibited a greater concordance rate (0.809) and Kappa coefficient (0.488), and outperformed the other methods in terms of the area under the ROC curve. Nanopore sequencing surpassed the Melting Curve method in identifying drug-resistant mutations.

Conclusion: Nanopore sequencing significantly enhances the detection of tuberculosis pathogens and drug-resistant genes.