ArticleWorld journal of microbiology & biotechnology2026
Comparative analysis of next-generation versus third-generation sequencing for pathogen detection in clinical samples: a diagnostic accuracy study.
Article in World journal of microbiology & biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundThe rapid and accurate identification of pathogens is crucial for clinical management of infectious diseases. While Next-generation sequencing (NGS) has transformed pathogen detection, Third-generation sequencing (TGS) offers advantages in real-time analysis and long-read capabilities. This study comprehensively compares the diagnostic performance of NGS and TGS across diverse clinical samples.
methodsWe conducted a prospective diagnostic accuracy study involving 105 clinical samples (58 bronchoalveolar lavage fluid, 28 whole blood, 19 other body fluids) from patients with suspected infections. All samples were analyzed using both NGS (BGI platform) and TGS (Nanopore platform). Diagnostic performance was evaluated against a composite reference standard incorporating clinical diagnosis, microbiological culture, and laboratory findings.
resultsNGS demonstrated significantly higher sensitivity compared to TGS (95.9% vs. 82.4%, p < 0.001), while TGS showed superior specificity (87.1% vs. 64.5%, p = 0.012). The overall agreement between platforms was 85.7% (Kappa = 0.702). NGS exhibited particular advantages in viral detection (32 vs. 8 detections, p < 0.001) and fungal identification (28 vs. 18 detections, p = 0.023), whereas both technologies showed comparable bacterial detection capabilities. The area under the ROC curve was 0.92 for NGS and 0.85 for TGS. Turnaround time was significantly shorter for TGS (median 8 h vs. 30 h, p < 0.001).
conclusionNGS and TGS demonstrate complementary strengths in clinical pathogen detection. NGS offers superior sensitivity and enhanced detection of viral and fungal pathogens, making it suitable for comprehensive diagnostic evaluation. TGS provides rapid results with higher specificity, advantageous for time-critical clinical decisions. A combined or scenario-specific approach may optimize pathogen detection in clinical practice.
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