石蜡组织中结核杆菌DNA检测在辅助诊断儿童结核病中的回顾性研究

A retrospective study on the detection of Mycobacterium tuberculosis DNA in paraffin tissues as an aid in the diagnosis of tuberculosis in children

目的探讨荧光探针PCR法检测石蜡组织中结核杆菌DNA在辅助诊断儿童结核病中的应用价值,同时分析、总结可能对实验结果产生影响的因素及注意事项。方法回顾性选取2017年8月至2019年9月重庆医科大学附属儿童医院用荧光探针PCR法检测石蜡组织中结核杆菌DNA的病例177例,将检测结果与石蜡切片抗酸染色及病理诊断结果进行比较,验证其诊断价值,同时总结荧光探针PCR法检测石蜡组织标本中结核杆菌的影响因素及注意事项。结果177例病例中84例诊断为结核、卡介苗病或符合结核、卡介苗病、结核伴感染,结核分枝杆菌DNA(荧光探针PCR法)检出阳性59例,荧光探针PCR法灵敏度为70.238%;其余93例病理诊断为肉芽性肿、化脓性炎症、猫抓病、淋巴瘤等非结核性疾病,结核分枝杆菌DNA检测结果均为阴性,荧光探针PCR法特异度为100%。151例同步行抗酸染色,荧光探针PCR法检出结核病灵敏度为69.333%(52/75),显著高于抗酸染色的14.667%(11/75),差异有统计学意义(P<0.05)。结论与结核杆菌抗酸染色比较,荧光探针PCR法检测组织中结核杆菌DNA具有更高的灵敏度,能显著提高结核病/卡介苗病的检出率,为临床诊疗提供有力证据。

Objective To explore the value of fluorescent probe PCR for detecting Mycobacterium tuberculosis DNA in paraffin tissues as an aid in the diagnosis of tuberculosis in children,and to analyze and summarize the factors that may have an impact on the experimental results and precautions at the same time.Methods A total of 177 cases for whom the Mycobacterium tuberculosis DNA in paraffin tissue had been detected by fluorescent probe PCR in the Children's Hospital of Chongqing Medical University from August 2017 to September 2019were retrospectively selected.The test results were compared with the acid-fast staining of paraffin sections and the pathological diagnosis or clinical results to validate the diagnostic value.Meanwhile,the influencing factors and precautions of fluorescent probe PCR for detecting Mycobacterium tuberculosis in paraffin tissue specimens were summarized.Results In the 177 cases,84 cases were diagnosed as tuberculosis,bacillus CalmetteGuérin or consistent with tuberculosis,bacillus Calmette-Guérin,tuberculosis with infection.Mycobacterium tuberculosis DNA(fluorescent probe PCR method)was detected positively in 59 cases,with a sensitivity rate of the fluorescent probe PCR method of 70.238%.The rest of the 93 cases were diagnosed with non-tuberculous diseases such as granulomas,suppurative inflammation,cat-scratch disease,lymphoma and so on.Their Mycobacterium tuberculosis DNA test results were negative,with the specificity of fluorescent probe PCR method of 100%.151 cases were treated with acid-fast staining simultaneously.The sensitivity of fluorescent probe PCR method for detecting tuberculosis was 69.333%(52/75),which was significantly higher than that of acid-fast staining at 14.667%(11/75),with a statistically significant difference(P<0.05).Conclusion Compared with Mycobacterium tuberculosis acid-fast staining,fluorescent probe PCR method has higher sensitivity in detecting Mycobacterium tuberculosis DNA in tissues,which can greatly improve the detection rate of tuberculosis/bacillus Calmette-Guérin disease,and provide strong evidence for clinical diagnosis and treatment.