Abstract:

Objective: To evaluate the application value of next generation sequencing (NGS) technology in the diagnosis of osteoarticular tuberculosis. Methods: A retrospective study method was used to collect the clinical data of 185 patients with suspected osteoarticular tuberculosis admitted to the Department of Orthopedics of Hebei Provincial Chest Hospital from Dec 2019 to Dec 2022 with reference to the enrollment criteria. The patients were categorized into the tuberculosis group (osteoarticular tuberculosis, 155 cases) and the non-tuberculosis group (non-osteoarticular tuberculosis, 30 cases) according to the final clinical diagnosis. All patients’ lesion specimens were obtained during surgery or puncture (including 51 pus, 89 granulation tissue and 45 bone tissue specimens), and all of them were sent for NGS, BACTEC MGIT 960 mycobacterial liquid culture (culture), and rifampicin resistance real-time fluorescence quantitative nucleic acid amplification assay (GeneXpert MTB/RIF, Xpert). The final clinical diagnosis was used as the reference standard to compare the efficacy of the three methods in the diagnosis of osteoarticular tuberculosis, and to analyze the positivity of the three methods in the detection of pus, granulation tissue and bone tissue specimens. Results: Of the 185 patients with suspected bony joint tuberculosis, the positive rate of NGS for detecting osteoarticular tuberculosis (63.24%, 117/185) was significantly higher than that of Xpert (54.05%, 100/185) and culture (35.68%, 66/185), and the differences were statistically significant (χ²=24.982, P<0.001; χ²=37.934, P<0.001). Using the final clinical diagnosis as the reference standard, the sensitivity of NGS, Xpert and culture methods for the diagnosis of osteoarticular tuberculosis were 74.84% (116/155), 64.52% (100/155) and 42.58% (66/155), respectively, and the specificity were 96.67% (29/30), 100.00% (30/30) and 100.00% (30/30), the diagnostic compliance rates were 78.38% (145/185), 70.27% (130/185) and 51.89% (96/185), respectively, and the Kappa values were 0.799, 0.590 and 0.504, respectively, and the AUC (95%CI) values were 0.867 (0.693-0.941), 0.703 (0.612-0.784) and 0.623 (0.529-0.717), respectively. The positive rates of pus detected by NGS was 80.39% (41/51), respectively, which were significantly higher than that granulation (62.92% (56/89)) and bone tissue specimens (44.44% (20/45)), and the differences were all statistically significant (χ²=4.560, P=0.031; χ²=13.335, P<0.001). Conclusion: NGS technology can significantly increase the pathogenetically positive detection rate in patients with osteoarticular tuberculosis with high detection efficacy, and has the highest diagnostic value with pus specimens.

Key words: Tuberculosis, osteoarticular, Metagenomic next-generation sequencing, Diagnostic techniques and procedures, Comparative study