高通量测序技术在脊柱感染病原体检测中的初步应用  

Application of high-throughput sequencing technology in pathogens detection for spinal infections

作　　者：刘富兵 王孝宾[1] 李晶[1] 胡嘉瑞[1] 张谦实 郑禹豪 王冰[1] 吕国华[1] Liu Fubing;Wang Xiaobin;Li Jing;Hu Jiarui;Zhang Qianshi;Zheng Yuhao;Wang Bing;Lyu Guohua(Department of Spine Surgery,the Second Xiangya Hospital of Central South University,Changsha 410011,China)

机构地区：[1]中南大学湘雅二医院脊柱外科,长沙410011

出　　处：《中华骨科杂志》2021年第3期149-156,共8页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金(81701228);湖南省自然科学基金(2018JJ3754)。

摘　　要：目的探讨高通量测序(high-throughput sequencing,HTS)技术在脊柱感染病原体检测中的应用价值。方法2019年1月至2020年5月收治疑似脊柱感染患者41例,男31例,女10例,年龄(59.7±11.9)岁(范围29~75岁)。有局部疼痛症状者37例、发热(≥38℃)15例、神经功能障18例。感染部位:颈椎4例,胸椎8例,腰椎29例。36例行病灶清除植骨融合内固定,5例行保守治疗(3例药物治疗,2例化疗)。开放手术获取的病灶组织送病理学检查和HTS检查。保守治疗的2例采用CT引导经皮穿刺获取组织块,1例采用B超引导经皮穿刺获取脓液,1例采集静脉血,1例行腰椎穿刺采集脑脊液。组织块送病理学检查和HTS检查,液体标本送细菌培养和HTS检查。以病理检查结果为"金标准",计算HTS检查的敏感度和特异度。以HTS结果为依据,结合临床表现、影像学检查和病理学检查结果,术后选择针对性的抗生素或抗结核药物治疗。细菌性感染患者术后抗感染治疗3个月;结核患者采用"四联"(异烟肼+利福平+吡嗪酰胺+乙胺丁醇)抗结核治疗1年。观察治疗前后血液炎症指标变化,监测病情进展及疗效。结果HTS病原体检测结果均在48 h内获得。初期的5例行一代测序验证,符合率为100%。41例中,26例结果阳性,阳性率63.4%,其中结核分枝杆菌13例(占31.7%)、葡萄球菌6例(占14.6%)、真菌和布鲁杆菌各2例(分别占4.9%)。15例检测结果为阴性,包括肿瘤或肿瘤样病变2例。38例行病理学检查,结果提示化脓性感染7例、结核12例、肿瘤或肿瘤样病变2例,其余17例为炎症性病变。HTS的敏感度为80.0%(16/20),特异度为55.6%(10/18),阳性预测值及阴性预测值分别为66.7%(16/24)和71.4%(10/14)。所有患者均随访3个月以上,治疗后3个月白细胞计数由入院时的(7.50±3.26)×10^9/L降至(6.22±2.53)×10^9/L,差异无统计学意义(t=1.082,P=0.290);C反应蛋白由(32.2±34.1)mg/L降至(4.5±10.5)mg/L,红细胞沉降率由(Objective To explore the application of high-throughput sequencing(HTS)technology in pathogens detection for spinal infection.Methods From January 2019 to May 2020,a total of 41 patients including 31 males and 10 females with an average age of 59.7±11.9 years(29-75 years)were suspected of spinal infections.There were 37 patients with local pain,15 with fever(≥38℃)and 18 with neurological dysfunction.The infected sites were as follows,4 cases of cervical spine,8 cases of thoracic spine and 29 cases of lumbar spine.There were 36 patients met the surgical indications and underwent open debridement,bone grafting,fusion and internal fixation,while the other 5 patients underwent conservative treatment(three received drug therapy and two were transferred to the internal department for chemotherapy).Lesions obtained from open surgery patients were underwent pathology and HTS examination.In 5 cases with conservative treatment,two of them underwent CT guided percutaneous puncture for samples,while one case underwent ultrasound guided percutaneous puncture for pus,one case for venous blood,and one case received lumbar puncture for cerebrospinal fluid.The samples were sent for pathological and HTS examination,while liquid specimens were sent for bacterial culture and HTS.The sensitivity and specificity of HTS results were determined according to pathological examination which was regarded as the"gold standard".Based on HTS results combined with the clinical manifestations,imaging examination and pathological results of the patients,targeted antibiotics or anti-tuberculosis drugs were selected for postoperative drug therapy.Patients with bacterial infection received anti-infection treatment for 3 months after operation.For tuberculosis patients,"tetrad"(isoniazid+rifampicin+pyrazinamide+ethambutanol)anti-tuberculosis treatments were underwent for one year.Inflammation indicators from the blood samples were observed before and after treatment,including white blood cell count(WBC),C-reactive protein(CRP)and erythrocyte sed

关 键 词：病原体检测 脊柱感染 化脓性感染 肿瘤样病变 腰椎穿刺 炎症性病变 病理学检查 白细胞计数 

分 类 号：R681.51[医药卫生—骨科学]