纳米孔靶向测序技术对福尔马林固定石蜡包埋组织中肉芽肿性疾病的诊断价值  

The value of nanopore-targeted sequencing technology in the diagnosis of granulomatous diseases in formalin-fixed paraffin-embedded tissues

作　　者：孙晓柯 魏金星[3] 张春艳[4] 梁瑞霞[2] 史慧敏[2] 阮祥林[2] 段鸿飞[1] Sun Xiaoke;Wei Jinxing;Zhang Chunyan;Liang Ruixia;Shi Huimin;Ruan Xianglin;Duan Hongfei(Department of Tuberculosis,Beijing Chest Hospital/Beijng Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149,China;Department of Tuberculosis,Henan Provincial Chest Hospital,Zhengzhou 450008,China;Department of Thoracic Surgery,Henan Provincial Chest Hospital,Zhengzhou 450008,China;Department of Respiratory and Critical Care Medicine,Henan Provincial Chest Hospital,Zhengzhou 450008,China)

机构地区：[1]首都医科大学附属北京胸科医院/北京市结核病胸部肿瘤研究所结核科,北京101149 [2]河南省胸科医院结核内科,郑州450008 [3]河南省胸科医院胸外科,郑州450008 [4]河南省胸科医院呼吸和危重症科,郑州450008

出　　处：《中国防痨杂志》2025年第5期582-588,共7页Chinese Journal of Antituberculosis

基　　金：河南省科技厅科技发展计划(232102310306)。

摘　　要：目的:评估纳米孔靶向测序技术(nanopore-targeted sequencing,NTS)对福尔马林固定石蜡包埋(formalin fixation and paraffin embedding,FFPE)组织中肉芽肿性疾病的检测能力。方法:参照入组标准收集2023年1—6月河南省胸科医院收治的因治疗需要行肺叶切除、淋巴结切除、骨关节手术治疗,且术后组织传统病理学结果提示为肉芽肿性炎的82例患者临床资料(包括传统病理学检测结果等),应用NTS技术检测术后病灶组织的FFPE标本,以最终临床诊断为参照评估NTS技术和传统病理学方法对肉芽肿性疾病的诊断价值。结果:82例患者中,组织病理学报告疑似结核病73例(89.0%)、结核分枝杆菌合并曲霉菌感染4例(4.9%)、隐球菌感染3例(3.7%)、肉芽肿性炎2例(2.4%);NTS检出菌种分别为结核分枝杆菌31例(37.8%)、真菌14例(17.1%)、非结核分枝杆菌9例(11.0%)、诺卡菌5例(6.1%)、布鲁杆菌和结核分枝杆菌合并黑曲霉各2例(2.4%)、未检出病原体16例(19.5%),以及结核分枝杆菌合并耶氏肺孢子菌、合并聚多曲霉、合并鸟分枝杆菌各1例(1.2%);最终临床诊断为结核病43例(52.4%)、真菌病13例(15.9%)、非结核分枝杆菌病9例(11.0%)、诺卡菌病和结核分枝杆菌合并真菌感染各5例(6.1%)、布鲁杆菌病2例(2.4%)、结核分枝杆菌合并非结核分枝杆菌1例(1.2%),4例(4.9%)患者诊断未明。以最终临床诊断为参照,病理学诊断对感染性肉芽肿的检测敏感度为60.3%(47/78),Kappa值为0.129;NTS检测感染性肉芽肿的敏感度为83.3%(65/78),Kappa值为0.328;组织病理学联合NTS技术检测感染性肉芽肿的敏感度为97.4%(76/78),Kappa值为0.788;三者特异度均为4/4。结论:NTS技术可检测肉芽肿性疾病到菌种水平,联合传统组织病理学可明显提高诊断感染性疾病病因学的效能,推荐作为传统组织病理学的补充方法。Objective:To evaluate the detection ability of nanopore-targeted sequencing(NTS)on granulomatous diseases in formalin-fixed paraffin-embedded(FFPE)tissues.Methods:According to inclusion criteria,clinical data of 82 patients admitted to Henan Provincial Chest Hospital from January to June 2023 who underwent lobectomy,lymph node dissection,and bone and joint surgical treatment due to therapeutic needs,and whose post-operative tissues were tested with traditional pathological tests which suggested granulomatous inflammation were collected(including the results of traditional pathological tests,etc.).NTS was applied on FFPE specimen from post-operative lesion tissue to compare its diagnostic value with traditional pathological methods on detecting granulomatous diseases,taking final clinical diagnosis as reference.Results:Of the 82 patients,histopathological test reported 73 cases(89.0%)of suspected tuberculosis,4 cases(4.9%)of tuberculosis combined with aspergillosis,3 cases(3.7%)of cryptococcosis,and 2 cases(2.4%)of granulomatous inflammation.The NTS detected 31 cases(37.8%)of Mycobacterium tuberculosis(MTB),14 cases(17.1%)of Fungi,9 cases(11.0%)of non-tuberculous mycobacterial(NTM),5 cases(6.1%)of Nocardia,2 cases(2.4%)of Brucella,2 cases(2.4%)of MTB combined with Aspergillus niger,16 cases(19.5%)of undetectable pathogens,and 1 case(1.2%)each of MTB combined with Pneumocystis carinii,Aspergillus polysporus,and M.avium.The final clinical diagnosis were 43 cases(52.4%)of tuberculosis,13 cases(15.9%)of fungal disease,9 cases(11.0%)of NTM disease,5 cases(6.1%)of each nocardiosis and tuberculosis in combination with fungal disease,2 cases(2.4%)of brucellosis,1 case(1.2%)of tuberculosis in combination with NTM disease,and 4 patients(4.9%)with an unspecified diagnosis.With reference to the final clinical diagnosis,the sensitivities of pathological diagnosis and NTS for the detection of granulomatous inflammation were 60.3%(47/78)and 83.3%(65/78),with a Kappa value of 0.129 and 0.328,respectively;and the sensitivity of co

关 键 词：肉芽肿病 慢性 石蜡包埋 组织固定 纳米技术 分子诊断技术 对比研究 

分 类 号：R52[医药卫生—内科学] R44[医药卫生—临床医学]