DataSheet_1_The diagnostic value of blood metagenomic next-generation sequencing in patients with acute hematogenous osteomyelitis.docx

AimsThis study aims to evaluate the diagnostic value of blood metagenomic next-generation sequencing (mNGS) in detecting pathogens from patients clinically diagnosed as acute hematogenous osteomyelitis (AHO).
MethodsThis retrospective study enrolled 66 patients with AHO. The test results of mNGS and bacterial culture on different samples, including blood and puncture fluid samples, from patients with AHO were compared to explore the diagnostic value of blood mNGS. Besides, this study also explored the efficacy of blood mNGS in decision making for antibiotic administration and analyzed the factors associated with the positive result of blood mNGS.
ResultsThe most common causative pathogens were Staphylococcus and Streptococcus. The sensitivity of blood mNGS (77.3%) was higher than that of blood culture (42.4%) (P<0.001), while the turnaround time of blood mNGS (2.1 ± 0.4 d) is much less than that of blood culture (6.0 ± 2.1 d) (P<0.001). Besides, the sensitivity of blood mNGS tests (77.3%) was slightly lower than that of puncture fluid mNGS (89.4%). Furthermore, detection comparison at pathogen level unravels that blood mNGS might be suitable for diagnosing AHO caused by common pathogens, while puncture fluid mNGS could be considered as preferred examination in diagnosing AHO caused by uncommon pathogens. Finally, three independent factors associated with the true positive result of blood mNGS in patients with AHO were identified, including Gram-positive pathogens (OR=24.4, 95% CI = 1.4-421.0 for Staphylococcus; OR=14.9, 95%CI= 1.6-136.1 for other Gram-positive bacteria), body temperature at sampling time (OR=8.2, 95% CI = 0.6-107.3 for body temperature of >38.5°C; OR=17.2, 95% CI = 2.0-149.1 for patients who were chilling), and no use of antibiotics before sampling (OR=8.9, 95% CI =1.4-59.0).
ConclusionThis is the first report on evaluating and emphasizing the importance of blood mNGS in diagnosing AHO. Blood sample might be an alternative sample for puncture fluid for mNGS, and its extensive application in diagnosing AHO could be expected.

研究目的 本研究旨在评估血液宏基因组二代测序（metagenomic next-generation sequencing, mNGS）对临床诊断为急性血源性骨髓炎（acute hematogenous osteomyelitis, AHO）患者的病原体检测诊断价值。
研究方法 本项回顾性研究纳入66例AHO患者。对比AHO患者血液、穿刺液等不同样本的mNGS与细菌培养检测结果，以探讨血液mNGS的诊断价值。此外，本研究还探索了血液mNGS在抗生素使用决策中的应用效能，并分析了血液mNGS检测结果呈阳性的相关影响因素。
研究结果 本研究检出的最常见致病病原体为葡萄球菌属与链球菌属。血液mNGS的诊断灵敏度为77.3%，高于血液培养的42.4%（P<0.001）；而血液mNGS的检测周转时间为2.1±0.4天，显著短于血液培养的6.0±2.1天（P<0.001）。此外，血液mNGS的灵敏度（77.3%）略低于穿刺液mNGS（89.4%）。进一步在病原体层面的检测对比显示，血液mNGS或适用于诊断常见病原体引发的AHO，而穿刺液mNGS可作为罕见病原体所致AHO的首选检测手段。最终本研究明确了3项与AHO患者血液mNGS真阳性结果相关的独立影响因素：包括革兰阳性病原体（葡萄球菌对应的比值比（odds ratio, OR）=24.4，95%置信区间（confidence interval, CI）=1.4~421.0；其他革兰阳性菌对应的OR=14.9，95%CI=1.6~136.1）、采样时的体温（体温>38.5℃对应的OR=8.2，95%CI=0.6~107.3；伴寒战患者对应的OR=17.2，95%CI=2.0~149.1），以及采样前未使用抗生素（OR=8.9，95%CI=1.4~59.0）。
研究结论 本研究为首项评估并强调血液mNGS在AHO诊断中重要价值的报道。血液样本可作为mNGS检测中穿刺液的替代样本，其在AHO诊断中的广泛应用值得期待。