Comparison of Xpert MTB/RIF Assay and the Targeted Next-Generation Sequencing for the Diagnosis of Pleural…

Cohort studyInfection and drug resistance· 2026

Comparison of Xpert MTB/RIF Assay and the Targeted Next-Generation Sequencing for the Diagnosis of Pleural Tuberculosis: A Prospective Comparative Diagnostic Accuracy Study.

Peng Wen, Shang Xue Wang, Xiao Man Ma … Min Wei

Molecular PathologyCohort studyQ2IF 3.2

tNGS found tuberculosis in 40% of culture-negative pleural cases vs 10% with Xpert

Comparison of Xpert MTB/RIF Assay and the Targeted Next-Generation Sequencing for the Diagnosis of Pleural Tuberculosis: A Prospective Comparative Diagnostic Accuracy Study.

Peng Wen … Min Wei

Infection and drug resistance · 2026

Purpose

To evaluate the diagnostic performance of targeted next-generation sequencing (tNGS), a multiplex PCR-based and hybrid-capture sequencing platform targeting 198 respiratory pathogens, versus Xpert MTB/RIF in diagnosing pleural tuberculosis (tuberculosis) across diverse specimen types, given the limited sensitivity of existing assays in paucibacillary disease.

Methods

Consecutive patients with suspected pleural tuberculosis were prospectively enrolled.

n = 105

40.0%

Results

40.0%

tNGS confirmed tuberculosis in 4x more culture-negative cases than Xpert did

n = 105

More results

In the overall cohort (N = 142), tNGS demonstrated higher sensitivity than Xpert MTB/RIF and mycobacterial culture across specimen types when evaluated against the composite reference standard, while maintaining 100% specificity in the non-tuberculosis group (n = 8).

More results

In sputum samples, tNGS achieved a sensitivity of 72.73%, significantly outperforming Xpert MTB/RIF (21.43%) and MTB culture (16.30%).

The paired analysis confirmed that tNGS significantly outperformed Xpert MTB/RIF in overall diagnostic yield ( p < 0.01), detecting 26 positive cases missed by Xpert.

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Conclusion · 1 of 2

tNGS provides incremental diagnostic value over conventional molecular and culture-based assays in pleural tuberculosis, particularly in paucibacillary and culture-negative disease.

Conclusion · 2 of 2

It increases microbiological confirmation in culture- and Xpert-negative cases and is best positioned as a complementary second-line assay within a tiered diagnostic framework, despite no assessment of downstream clinical impact.