基于病理组织行RNA原位杂交技术联合原位免疫荧光染色检测结核分枝杆菌的应用研究

姚利利; 欧玉荣; 吴礼高

doi:10.13898/j.cnki.issn.1000-2200.2024.12.018

基于病理组织行RNA原位杂交技术联合原位免疫荧光染色检测结核分枝杆菌的应用研究

Application value of the RNA in situ hybridization combined with in situ immunofluorescence staining based on pathological tissue in the detection of Mycobacterium tuberculosis

摘要

摘要: 目的：探讨RNA原位杂交技术联合原位免疫荧光染色在结核分枝杆菌检测中的应用价值。方法：采用回顾性研究方法，选取肺或淋巴结穿刺或者手术标本病理诊断为肉芽肿性炎病人100例作为研究对象。其中70 例为确诊结核病病人(确诊组)，30例为非结核病病人(对照组)。以实时荧光定量PCR检测结果为参照，均进行抗酸染色、原位免疫荧光染色、RNA原位杂交技术检测。评价RNA原位杂交技术联合原位免疫荧光染色检测结核分枝杆菌的诊断效能。结果：以实时荧光定量PCR检测结果为金标准，抗酸染色的灵敏度、特异度、阳性预测值、阴性预测值、准确度、约登指数分别为30.00%、93.33%、91.33%、36.36%、49.00%、0.233，原位免疫荧光染色分别为51.43%、93.33%、94.74%、45.16%、64.00%、0.447，RNA原位杂交技术分别为77.14%、100.00%、100.00%、65.22%、84.00%、0.771，RNA原位杂交技术联合原位免疫荧光染色分别为84.29%、100.00%、100.00%、73.17%、89.00%、0.843。RNA原位杂交技术联合原位免疫荧光染色的ROC曲线下面积(AUC)为0.951，RNA原位杂交技术AUC为0.768，原位免疫荧光染色AUC为0.843。结论：RNA原位杂交技术联合原位免疫荧光染色的灵敏度及特异度、阳性预测值、阴性预测值、准确度及约登指数均较高，是一种具有潜力的结核分枝杆菌检测方法。

Abstract: Objective: To explore the application value of in situ immunofluorescence staining combined with RNA in situ hybridization technology in the detection of Mycobacterium tuberculosis. Methods: A retrospective study method was used to select 100 granulomatous inflammation patients with lung or lymph node puncture or surgical specimen diagnosed by pathological biopsy as the study subjects.Of these,70 cases were confirmed tuberculosl(TB) patients(confirmed group),and 30 cases were non-TB patients(control group).Based on the results of real-time fluorescence quantitative PCR,the acid-fast staining,in situ immunofluorescence staining and RNA in situ hybridization were performed.The diagnostic efficacy of RNA in situ hybridization combined with in situ immunofluorescence staining in the detection of Mycobacterium tuberculosis was evaluated. Results: Taking the results of real-time fluorescence quantitative PCR as the gold standard,the sensitivity,specificity,positive predictive value,negative predictive value,accuracy and Youden index of acid-fast staining were 30.00%,93.33%,91.33%,36.36%,49.00% and 0.233,respectively;the sensitivity,specificity,positive predictive value,negative predictive value,accuracy and Youden index of in situ immunofluorescence staining were 51.43%,93.33%,94.74%,45.16%,64.00% and 0.447,respectively;the sensitivity,specificity,positive predictive value,negative predictive value,accuracy and Youden index of RNA in situ hybridization were 77.14%,100.00%,100.00%,65.22%,84.00% and 0.771,respectively;the sensitivity,specificity,positive predictive value,negative predictive value,accuracy and Youden index of in situ immunofluorescence staining combined with RNA in situ hybridization were 84.29%,100.00%,100.00%,73.17%,89.00% and 0.843,respectively.The ROC curve AUC value of in situ immunofluorescence staining combined with RNA in situ hybridization technology was 0.951,the ROC curve AUC value of RNA in situ hybridization technology was 0.768,and the ROC curve AUC value of in situ immunofluorescence staining was 0.843. Conclusions: The sensitivity and specificity,positive predictive value,negative predictive value,accuracy and Youden index of in situ immunofluorescence staining combined with RNA in situ hybridization technology are high.It is a potential method for detecting Mycobacterium tuberculosis.

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