CRISPR-Cas13a检测在肺结核诊断中的价值  

Value of CRISPR-Cas13a detection in the diagnosis of pulmonary tuberculosis

作　　者：张治国[1] 尚媛媛[2] 张旭霞[2] 刘荣梅 马丽萍[4] 秦林[5] 孔忠顺[3] 任卫聪[2] Zhang Zhiguo;Shang Yuanyuan;Zhang Xuxia;Liu Rongmei;Ma Liping;Qin Lin;Kong Zhongshun;Ren Weicong(Department of Tuberculosis,Changping District for Tuberculosis Research and Control,Beijing 102200,China;Department of Bacteriology and Immunology,Beijing Chest Hospital,Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149,China;DepartmentⅡof Tuberculosis,Beijing Chest Hospital,Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149,China;DepartmentⅠof Tuberculosis,Beijing Chest Hospital,Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149,China;Department of Endoscopic Diagnosis and Treatment,Beijing Chest Hospital,Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing 101149,China)

机构地区：[1]北京市昌平区结核病防治所,北京102200 [2]北京市结核病胸部肿瘤研究所/首都医科大学附属北京胸科医院耐药结核病研究北京市重点实验室细菌免疫室,北京101149 [3]北京市结核病胸部肿瘤研究所/首都医科大学附属北京胸科医院结核二科,北京101149 [4]北京市结核病胸部肿瘤研究所/首都医科大学附属北京胸科医院结核一科,北京101149 [5]北京市结核病胸部肿瘤研究所/首都医科大学附属北京胸科医院内镜诊疗科,北京101149

出　　处：《中国防痨杂志》2023年第6期589-593,共5页Chinese Journal of Antituberculosis

基　　金：“十三五”国家科技重大专项(2018ZX10301407-006);首都卫生发展科研专项(2020-1-1041)。

摘　　要：目的:评价应用CRISPR-Cas13a检测临床样本对肺结核的诊断价值。方法:采用回顾性研究方法,收集首都医科大学附属北京胸科医院2022年9月至2022年12月收治住院的102例经GeneXpert MTB/RIF检测(简称“Xpert检测”)确诊的肺结核患者的支气管肺泡灌洗液标本,作为结核病组;收集同期收治入院的100例非结核病患者(包括肺癌,细菌性肺炎,支气管扩张,慢性阻塞性肺疾病)的支气管肺泡灌洗液标本,作为非结核病组。所有患者的支气管肺泡灌洗液标本均进行涂片抗酸杆菌染色镜检、培养、Xpert检测及CRISPR-Cas13a检测。以Xpert检测结果为参照,评价CRISPR-Cas13a检测支气管肺泡灌洗液对肺结核的诊断价值。结果:结核病组102份标本中,CRISPR-Cas13a检测阳性为99份(97.1%),涂片阳性40份(39.2%),培养阳性68份(66.7%)。CRISPR-Cas13a检测阳性率明显高于涂片和培养法,差异有统计学意义(涂片:χ^(2)=58.141,P<0.001;培养:χ^(2)=30.250,P<0.001)。与传统检测方法相比,CRISPR-Cas13a比涂片和培养法多检测出28份标本。Xpert检测结核病组阳性率为100.0%;Xpert检测非结核病组阴性率为100.0%。以Xpert检测结果作为参照,CRISPR-Cas13a检测肺结核的敏感度为97.1%(99/102)、特异度为96.0%(96/100),准确度为96.5%(195/202),阳性预测值为96.1%(99/103),阴性预测值为97.0%(96/99)。结论:CRISPR-Cas13a检测与传统方法相比简单、快捷,检测支气管肺泡灌洗液对肺结核的诊断具有较高敏感度及特异度,具有良好的应用前景。Objective:To assess the diagnostic value of CRISPR-Cas13a detection in pulmonary tuberculosis(TB)using clinical samples.Methods:A retrospective study was conducted at Beijing Chest Hospital Affiliated to Capital Medical University.The bronchoalveolar lavage fluid(BALF)specimens of 102 patients with pulmonary TB confirmed by GeneXpert MTB/RIF test(Xpert test)from September 2022 to December 2022 were enrolled as TB group,and the BALF specimens of 100 patients(including lung cancer,bacterial pneumonia,bronchiectasis,and chronic obstructive pulmonary disease)during the same period were recruited as non-TB group.The BALF samples of all patients were subjected to acid-fast bacilli smear microscopy,culture,Xpert test and CRISPR-Cas13a test.The diagnostic value of CRISPR-Cas13a in the detection of BALF for pulmonary TB was evaluated based on the results of Xpert test.Results:Among 102 specimens in the TB group,99(97.1%)were positive for CRISPR-Cas13a,40(39.2%)were positive for smear,and 68(66.7%)were positive for culture.The positive detection rate of CRISPR-Cas13a was significantly higher than those of smear and culture(smear:χ^(2)=58.141,P<0.001;culture:χ^(2)=30.250,P<0.001).Compared with traditional methods,CRISPR-Cas13a was able to identify 28 clinically confirmed TB patients which would be otherwise missed by either smear or culture.The positive detection rate of Xpert was 100.0%in TB group,and the negative detection rate of Xpert was 100.0%in non-TB group.Taking the results of Xpert as a reference,the sensitivity,specificity,accuracy,positive predictive value and negative predictive value of CRISPR-Cas13a in detecting pulmonary TB were 97.1%(99/102),96.0%(96/100),96.5%(195/202),96.1%(99/103)and 97.0%(96/99),respectively.Conclusion:Compared with traditional tests,CRISPR-Cas13a detection is simple and fast,with higher sensitivity and higher specificity in the detection of BALF for the diagnosis of pulmonary TB,and has a good application prospect.

关 键 词：重复序列 核酸 结核 肺 支气管肺泡灌洗液 CRISPR-Cas系统 

分 类 号：R521[医药卫生—内科学] R8[医药卫生—临床医学]