三种分子诊断技术对儿童肺结核的诊断价值

目的 探讨三种分子诊断技术对儿童肺结核的诊断效能,为儿童肺结核的诊断寻求简单、快速、准确的新方法。方法 搜集哈尔滨市胸科医院儿童结核科、哈尔滨市儿童医院呼吸科、哈尔滨医科大学附属第一医院儿科2016年9月至2018年6月收治的2个月至14周岁的疑似肺结核患儿共186例,最后临床诊断肺结核者119例,其中低龄儿54例(45.4%),年长儿65例(54.6%);非结核病者67例,其中低龄儿31例(42.3%),年长儿36例(53.7%)。应用实时荧光定量PCR(real-time fluorescen quantitative PCR,FQ-PCR)、实时荧光核酸恒温扩增检测技术(simultaneous amplification and testing,SAT)和耐药基因芯片技术分别检测患儿胃液和痰液(年长儿)中的结核分枝杆菌,评价各方法的诊断效能。结果 以临床诊断为标准,FQ-PCR、SAT、耐药基因芯片检测胃液中MTB的敏感度分别为81.51%(97/119)、 78.15%(93/119)、73.11%(87/119),特异度分别为86.57%(58/67)、98.51%(66/67)、92.54%(62/67),Kappa值分别为 0.653、0.709、0.603。3种方法检测年长儿胃液中MTB的敏感度分别为 80.00%(52/65)、73.85%(48/65)、67.69%(44/65),特异度分别为75.00%(27/36)、97.22%(35/36)、88.89%(32/36);检测年长儿痰液中MTB的敏感度分别为 47.69%(31/65)、41.54%(27/65)、36.92%(24/65),特异度分别为88.89%(32/36)、 97.22%(35/36)、91.67%(33/36),各方法检测的敏感度胃液均高于痰液,差异均有统计学意义(χ ²值分别为14.696、13.898、12.334,P值均为0.000),各方法检测的特异度胃液和痰液之间差异均无统计学意义(χ ²值分别为2.347、0.000、0.158,P值分别为0.126、1.000、0.691);胃液总的阳性检出率为87.69%(57/65),痰液总的阳性检出率为58.46%(38/65),两者差异有统计学意义(χ ²=14.114,P=0.000)。 结论 FQ-PCR、SAT、耐药基因芯片检测儿童疑似肺结核患者的胃液和痰液中结核分枝杆菌的敏感度高、特异度强,对儿童肺结核的诊断具有重要价值。

Diagnostic value of three molecular techniques for children pulmonary tuberculosis

Objective To investigate the diagnostic efficacy of three molecular diagnostic techniques for children pulmonary tuberculosis in order to find a simple, rapid and accurate method for the diagnosis of tuberculosis in children.Methods A total of 186 children from 2 months to 14 years old with suspected pulmonary tuberculosis who were admitted to the Children’s Tuberculosis Department of Harbin Chest Hospital, Respiratory Department of Harbin Children’s Hospital, Department of Pediatrics, the First Affiliated Hospital of Harbin Medical University from September 2016 to June 2018. Finally 119 cases were diagnosed as tuberculosis, including 54 cases (45.4%) of younger children, 65 cases (54.6%) of elderly children, compared with 31 cases (42.3%) of younger children and 36 cases (53.7%) of elderly children in 67 non-tuberculosis children. Real-time fluorescence quantitative PCR (FQ-PCR), simultaneous amplification and testing (SAT) and drug-resistant gene chip technology were used to detect Mycobacterium tuberculosis in gastric juice and sputum (older children) respectively to observe the diagnostic efficacy of each method.Results According to the clinical diagnosis, the sensitivity of FQ-PCR, SAT and drug-resistant gene chip detection of gastric juice were 81.51% (97/119), 78.15% (93/119), 73.11% (87/119), and the specificity were 86.57% (58/67), 98.51% (66/67), 92.54% (62/67) respectively, Kappa values were 0.653, 0.709, 0.603. The sensitivity of the three methods for detecting gastric juice in elderly children was 80.00% (52/65), 73.85% (48/65) and 67.69% (44/65), and the specificity was 75.00% (27/36), 97.22%(35/36) and 88.89% (32/36) respectively. The sensitivity of detecting sputum in elderly children was 47.69% (31/65), 41.54% (27/65), 36.92% (24/65), and the specificity was 88.89% (32/36), 97.22% (35/36) and 91.67% (33/36) respectively. The sensitivity of gastric juice detected by each method was higher than that of sputum, and the difference was statistically significant (χ ² values were 14.696, 13.898, 12.334, P values were 0.000). The specificity of each detection method was not statistical significance between gastric juice and sputum (χ ² values were 2.347, 0.000, 0.158, respectively, P values were 0.126, 1.000, 0.691). The total positive rate of gastric juice was 87.69% (57/65), and the sputum was 58.46% (38/65), the difference was statistically significant (χ ²=14.114, P=0.000). Conclusion FQ-PCR, SAT and drug-resistant gene chip detection methods have high sensitivity and specificity in the detection of Mycobacterium tuberculosis in gastric juice and sputum of children with suspected pulmonary tuberculosis, which is of great value in the diagnosis of tuberculosis in children.