A diagnostic performance study of multiplex polymerase chain reaction-based targeted next-generation sequencing for the accurate identification of Mycobacterium tuberculosis in bronchoalveolar lavage fluid
Abstract
Objective: To evaluate the diagnostic performance of multiplex PCR-based targeted next-generation sequencing (mp-tNGS) for detecting Mycobacterium tuberculosis (MTB) in bronchoalveolar lavage fluid (BALF) from patients with suspected pulmonary tuberculosis (PTB).
Methods: This prospective study enrolled 188 patients. BALF samples were tested in parallel by AFB smear, culture, Xpert MTB/RIF, and mp-tNGS. Diagnostic performance was assessed against a composite clinical reference standard; phenotypic drug susceptibility testing (pDST) was additionally compared in a subset of 35 patients.
Results: Among the cohort, 116 patients were diagnosed with PTB. The sensitivities of AFB smear, culture, Xpert MTB/RIF, and mp-tNGS were 19.8%, 33.6%, 73.3%, and 83.6%, respectively; specificities were 91.7%, 100%, 98.6%, and 97.2%. mp-tNGS demonstrated the highest diagnostic accuracy (88.8%) and the strongest agreement with the reference standard (κ = 0.868, p rpoB S450L mutations) and 75% sensitivity with 100% specificity for isoniazid resistance (katG S315T mutations), identifying five probable multidrug-resistant cases. Xpert MTB/RIF detected rifampicin resistance with 75.0% sensitivity and 96.3% specificity.
Conclusion: The mp-tNGS assay provides a superior combination of rapid turnaround time (~12 hours) and high diagnostic accuracy for MTB detection in BALF. Beyond pathogen identification, it offers rapid and preliminary genotypic resistance screening, supporting early suspicion of…
How strong is the evidence?
Evidence Hub reads this as a cohort study, done in people. Its evidence level is 3 of 5 (Moderate · trial / cohort / secondary analysis). Trials without a large randomized design, cohorts and secondary analyses give moderate evidence. Reported size: 188 participants or records.
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