Evaluation of the value of imaging combined with molecular diagnostic techniques in the diagnosis of tuberculosis
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摘要:
目的评估GeneXpert MTB/RIF、TaqMan探针荧光定量PCR、RNA恒温扩增法3种不同原理的分子生物学检验方法联合影像学表现对结核病诊断的价值。 方法收集胸部X线检查表现怀疑结核分枝杆菌感染的患者标本38例,以临床诊断结核病为金标准,比较GeneXpert MTB/RIF、TaqMan探针荧光定量PCR法、RNA恒温扩增法3种分子生物学检验方法的敏感度、特异性、阳性预测值和阴性预测值。 结果胸部X线检查怀疑结核分枝杆菌感染的患者中,23名临床诊断为结核病,15名临床排除结核病。GeneXpert MTB/RIF、TaqMan探针荧光定量PCR、RNA恒温扩增法检测的敏感度分别为87.0%、52.2%和13.0%,阴性预测值分别为83.3%、57.7%和43.2%,差异具有统计学意义(P < 0.05)。3种方法的阳性预测值均为100%。GeneXpert MTB/RIF方法的敏感度和阴性预测值最高,其次为TaqMan探针荧光定量PCR法,RNA恒温扩增法敏感度和阴性预测值最低。 结论对于胸部X线检查怀疑结核分枝杆菌感染的患者,GeneXpert MTB/RIF检测结核分枝杆菌具有更高的敏感度,在结核病诊断中具有较好的辅助诊断价值,应推荐成为结核分枝杆菌感染检测的首选分子生物学方法。 -
关键词:
- 结核分枝杆菌 /
- 胸部X线 /
- GeneXpert MTB/RIF /
- TaqMan探针荧光定量PCR /
- RNA恒温扩增
Abstract:ObjectiveTo evaluate the diagnostic value of three different molecular methods, GeneXpert MTB/RIF, TaqMan probe fluorescence quantitative PCR, and RNA constant temperature amplification, combined with imaging findings in the diagnosis of tuberculosis. MethodsWe collected 38 patient of suspected Mycobacterium tuberculosis infection on chest X-ray. The sensitivity, specificity, positive predictive value and negative predictive value of the three different molecular methods were compared. GeneXpert MTB/RIF, TaqMan probe fluorescence quantitative PCR, and RNA constant temperature amplification, when the clinical diagnosis of tuberculosis was taken as the gold standard. ResultsAmong the patients whose imaging findings were suspected of Mycobacterium tuberculosis infection, 23 were clinically diagnosed with tuberculosis, and 15 were clinically excluded from tuberculosis. The sensitivity of GeneXpert MTB/RIF, TaqMan probe fluorescence quantitative PCR, and RNA constant temperature amplification were 87.0%, 52.2% and 13.0%, respectively. The negative predictive value were 83.3%, 57.7% and 43.2%, respectively (P < 0.05). The positive predictive value of the three methods is 100%.GeneXpert MTB/RIF method had the highest sensitivity and negative predictive value, followed by TaqMan probe fluorescence quantitative PCR, and RNA isothermal amplification had the lowest sensitivity and negative predictive value. ConclusionFor patients with suspected imaging findings of Mycobacterium tuberculosis infection, GeneXpert MTB/RIF has higher sensitivity in detecting Mycobacterium tuberculosis, and good auxiliary diagnostic value in the diagnosis of tuberculosis. It should be recommended as a test for Mycobacterium tuberculosis infection. -
表 1 38例标本情况
Table 1. Types of 38 specimens(n)
项目 肺泡灌洗液 咳痰 胸水 腹水 穿刺液 临床诊断(金标准) 结核标本 21 0 0 1 1 23 排除结核标本 4 8 2 0 1 15 合计 25 8 2 1 2 38 表 2 3种方法对TB感染检出率比较
Table 2. Comparison of the detection rate of TB infection by three methods(n)
分类 检测方法 GeneXpert MTB/RIF TaqMan探针荧光定量PCR RNA恒温扩增法 检出TB 20 12 3 未检出TB 18 26 35 检出率(%) 52.6 31.6 7.9 表 3 GeneXpert MTB/RIF、TaqMan探针荧光定量PCR及RNA恒温扩增性能比较
Table 3. Comparison of the results of GeneXpert MTB/RIF, TaqMan probe fluorescence quantitative PCR and RNA constant temperature amplification(%)
检测方法 敏感度 特异性 阳性预测值 阴性预测值 GeneXpert MTB/RIF 87.00 100.00 100.00 83.30 TaqMan探针荧光定量PCR 52.20 100.00 100.00 57.70 RNA恒温扩增 13.00 100.00 100.00 43.20 -
[1] Chakaya J, Khan M, Ntoumi F, et al. Global Tuberculosis Report 2020 - Reflections on the Global TB burden, treatment and prevention efforts[J]. Int J Infect Dis, 2021 http://www.sciencedirect.com/science/article/pii/S1201971221001934 [2] Kanchar A, Swaminathan S. Tuberculosis control: WHO perspective and guidelines[J]. Indian J Pediatr, 2019, 86(8): 703-6. doi: 10.1007/s12098-019-02989-2 [3] Wang HY, Lu JJ, Chang CY, et al. Development of a high sensitivity TaqMan-based PCR assay for the specific detection of Mycobacterium tuberculosis complex in both pulmonary and extrapulmonary specimens[J]. Sci Rep, 2019, 9(1): 113. doi: 10.1038/s41598-018-33804-1 [4] Sharma G, Tewari R, Dhatwalia SK, et al. A loop-mediated isothermal amplification assay for the diagnosis of pulmonary tuberculosis[J]. Lett Appl Microbiol, 2019, 68(3): 219-25. doi: 10.1111/lam.13115 [5] 张正冬, 张海燕, 林存智. WHO第四版结核病治疗指南解读[J]. 中华临床医师杂志: 电子版, 2014, 8(23): 4251-3. https://www.cnki.com.cn/Article/CJFDTOTAL-ZLYD201423019.htm [6] Wu G, Fan Q. Pancreatic tuberculosis: a clinical diagnostic challenge [J]. ANZ J Surg, 2021, 91(4): 747-9. doi: 10.1111/ans.16249 [7] 张培元. 肺结核诊断和治疗指南[J]. 中华结核和呼吸杂志, 2001, 24 (2): 70-4. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHJH200204029.htm [8] 李忠奇, 吴冰心, 刘巧, 等. 结核病实验室诊断技术研究进展[J]. 中国热带医学, 2020, 20(4): 381-4. https://www.cnki.com.cn/Article/CJFDTOTAL-RDYX202004022.htm [9] 刘旭晖, 张爱梅, 卢水华. Xpert MTB/RIF试验用于肺结核快速检测的临床研究[J]. 中华肺部疾病杂志: 电子版, 2016, 9(5): 524-7. doi: 10.3877/cma.j.issn.1674-6902.2016.05.012 [10] Antonenka U, Hofmann-Thiel S, Turaev L, et al. Comparison of Xpert MTB/RIF with ProbeTec ET DTB and COBAS TaqMan MTB for direct detection of M. tuberculosis complex in respiratory specimens[J]. BMC Infect Dis, 2013, 13: 280. doi: 10.1186/1471-2334-13-280 [11] 刘宁, 付洪义, 孙金昊, 等. 结核分枝杆菌RNA恒温扩增实时检测法诊断活动性肺结核分析[J]. 中国预防医学杂志, 2019, 20(9): 789-92. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYC201909006.htm [12] Mboneni TA, Eales OO, Mosina NL, et al. Molecular detection of Mycobacterium tuberculosis in poor-quality cough specimens[J]. J Med Microbiol, 2020, 69(9): 1179-82. doi: 10.1099/jmm.0.001239 [13] 黄芳, 党丽云, 孙惠平, 等. 三种分子生物学诊断技术对结核病诊断价值的比较[J]. 中华结核和呼吸杂志, 2015, 38(9): 680-5. doi: 10.3760/cma.j.issn.1001-0939.2015.09.013 [14] 杜鹏, 郑文斌, 卢留珠, 等. Xpert MTB/RIF和荧光定量PCR在痰液标本中快速检测结核分支杆菌的比较[J]. 中国实验诊断学, 2018, 22 (4): 656-9. doi: 10.3969/j.issn.1007-4287.2018.04.032 [15] Steingart KR, Sohn H, Schiller I, et al. Xpert® MTB/RIF assay for pulmonary tuberculosis and rifampicin resistance in adults[J]. Cochrane Database Syst Rev, 2013(1): CD009593. [16] 徐东芳, 王庆. Xpert MTB/RIF系统在结核分枝杆菌和利福平耐药性快速检测中的应用[J]. 检验医学, 2017, 32(8): 722-6. doi: 10.3969/j.issn.1673-8640.2017.08.014 [17] Morel F, Jaffré J, Sougakoff W, et al. Place de la biologie moléculaire dans le diagnostic de la tuberculose[J]. Revue Des Maladies Respir, 2020, 37(5): 412-6. doi: 10.1016/j.rmr.2019.09.004