AUTHOR=Tao Zhengyong , Zhu Mengqi , Shi Jiandang , Yang Zongqiang , Niu NingKui TITLE=The application value of targeted next-generation sequencing in the diagnosis of primary osteoarticular infections: A single-center study JOURNAL=Frontiers in Cellular and Infection Microbiology VOLUME=Volume 15 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1593228 DOI=10.3389/fcimb.2025.1593228 ISSN=2235-2988 ABSTRACT=ObjectiveTo evaluate the diagnostic performance and clinical utility of targeted next-generation sequencing (tNGS) in primary osteoarticular infections (POI).MethodsEighty-seven patients diagnosed with POI at the Bone Infection Ward of Ningxia Medical University General Hospital between September 2023 and September 2024 were enrolled, including cases of tuberculous osteoarticular infection (35 cases), Brucella-related osteoarticular infection (21 cases), and pyogenic osteoarticular infection (31 cases). Using bacterial culture, Xpert MTB/RIF assay, Brucella agglutination test, and histopathological examination as reference standards, the diagnostic value of tNGS in pathogen identification and resistance gene analysis was systematically evaluated.ResultsAll patients had complete follow-up data. The cohort comprised 87 POI patients (mean age: 55.36 ± 17.24 years; male-to-female ratio: 1.35:1). tNGS demonstrated significantly higher overall sensitivity than conventional bacterial culture (85.0% vs. 31.0%, P < 0.001). For resistance profiling, tNGS identified Mycobacterium tuberculosis complex mutations associated with resistance to isoniazid (2 cases), rifampicin (2 cases), ethambutol (1 case), pyrazinamide (5 cases), and streptomycin (1 case). Additionally, one fluoroquinolone resistance gene and one extended-spectrum β-lactamase (ESBL)-producing pathogen were detected. Notably, one multidrug-resistant (MDR) case harbored mutations conferring resistance to five anti-tuberculosis agents. Receiver operating characteristic (ROC) curve analysis revealed that tNGS exhibited superior diagnostic accuracy for tuberculous osteoarticular infections (AUC = 0.926), Brucella-related osteoarticular infections (AUC = 0.891), and pyogenic osteoarticular infections (AUC = 0.912), outperforming Xpert MTB/RIF (0.814), Brucella agglutination test (0.832), bacterial culture (0.652), and histopathology (0.704) (all P < 0.05).ConclusiontNGS enables simultaneous pathogen identification and resistance gene detection with high efficiency, broad coverage, and accuracy, demonstrating significant advantages in POI diagnosis. This technology holds critical value in guiding optimized antimicrobial therapy and is recommended as a first-line molecular diagnostic tool for POI.