宏基因组二代测序技术在脊柱感染病原微生物诊断中的应用

Pathogenic detection by metagenomic next-generation sequencing in spinal infections

中文关键词:  脊柱感染  宏基因组二代测序  病原微生物

英文关键词:Spinal infection  Metagenomic  Next-generation sequencing  Pathogens

中文摘要:

  【摘要】 目的：探讨宏基因组二代测序（metagenomic next-generation sequencing，mNGS）技术在脊柱感染疾病中诊断病原微生物的能力和价值。方法：纳入2019年1月～2020年12月收治的46例疑似脊柱感染患者，其中男性26例，女性20例；年龄26～77岁（50.4±15.7岁）。获取患者的血液和病灶组织或脓液标本（CT引导下穿刺活检21例，开放手术25例），行微生物培养、血清学检测、病理检查和mNGS检测后进行结果的比较分析。结果：46例标本均在48h内获得了mNGS结果，微生物培养、血清学检测、病理检查等传统实验室检测时间1~12d不等。经临床最终诊断共35例为脊柱感染，11例为非脊柱感染。35例脊柱感染患者中，化脓性细菌感染14例，结核感染5例，布鲁菌属感染4例，未明确病原微生物感染12例；11例非脊柱感染患者中，脊柱肿瘤3例，终板Modic改变4例，终板骨折1例，DISH病合并假关节1例，强直性脊柱炎合并假关节2例。通过mNGS检测发现了脆弱拟杆菌、微小微单胞菌、齿垢密螺旋体、贝纳柯克斯体等少见病原微生物。mNGS在种水平与临床最终诊断的一致性为82.61%（19/23），在属水平的一致性为95.65%（22/23）。微生物培养阳性率为42.86%（15/35），mNGS检测阳性率为94.29%（33/35），二者存在统计学差异（P＜0.01）。mNGS检测的敏感度为91.43%，特异度为90.91%，阳性预测值96.97%，阴性预测值76.92%。结论：宏基因组二代测序技术检测病原微生物快速、高效、准确，在脊柱感染疾病的诊疗中有较高的诊断价值。

英文摘要:

  【Abstract】 Objectives: To evaluate the application and value of metagenomic next-generation sequencing (mNGS) in detecting pathogens from spinal infections. Methods: 46 samples were prospectively collected from patients who were suspected spinal infections and treated from January 2019 to December 2020. There were 26 males and 20 females, with an average age of 50.4±15.7 years(26-77 years). Blood and focal tissue or pus samples were obtained(21 samples were from CT guided biopsy while 25 samples from open surgery). Microbial culture, serological testing, pathological examination and mNGS were performed and compared pairwise in all cases. Results: All results of mNGS were obtained within 48 hours, and the traditional laboratory tests ranged from 1 to 12 days. Of all 46 cases, 35 were diagnosed as spinal infection and 11 cases as non-infection. 23 cases were found with microbiology evidence, including pyogenic infection(14 cases), tuberculosis(5 cases) and brucella(4 cases) infection. Still, 12 cases were clinically diagnosed as spinal infection without clear microbiology evidence. 11 cases were non-infection, including spine tumor(3 cases), endplate Modic change(4 cases), endplate fracture(1 case), Dish disease with pseudoarthrosis(1 case) and ankylosing spondylitis with pseudoarthrosis(2 cases). Rare microorganisms such as Bacteroides fragilis, Parvimonas micra, Treponema denticola and Coxiella burnetii were detected by mNGS. At the species level, the consistency between mNGS and the final clinical diagnosis was 82.61%(19/23), while that of mNGS reached 95.65%(22/23) at the genus level. The positive rate of mNGS was 94.29%(33/35), significantly higher than that of microbiological culture, which had a positive rate of 42.86%(15/35), P<0.01. The sensitivity, specificity, positive predictive value and negative predictive value of mNGS were 91.43%, 90.91%, 96.97% and 76.92%, respectively. Conclusions: mNGS is a fast, efficient and accurate diagnostic tool for pathogenic detection. It has high diagnostic and application value in the diagnosis and treatment of spinal infection diseases.