萋–尼氏抗酸染色、金胺“O”荧光染色、结核分枝杆菌DNA及培养法在结核分枝杆菌检测中的应用比较

Comparison of Ziehl-Neelsen Acid-Fast Staining, Auramine “O” Fluorescence Staining, Mycobacterium tuberculosis DNA Detection and Culture Method in the Detection of Mycobacterium tuberculosis

摘要: 目的：对比分析萋–尼氏抗酸染色、“O”荧光染色、结核分枝杆菌DNA检测及固体培养法四种检测方式对结核分枝杆菌的诊断效能，为结核病早期诊断提供参考依据。方法：选取218份痰液或支气管/肺泡灌洗液样本，分别采用四种方法开展检测，统计各组阳性检出率，比较不同检测方法的灵敏度、特异度、阳性预测值、阴性预测值，采用韦恩图分析阳性样本分布特征。结果：结核分枝杆菌DNA检测阳性检出率为22.02%，显著高于其余三种传统检测方法(P < 0.01)。诊断效能结果显示，以临床诊断为标准，结核分枝杆菌DNA检测灵敏度97.62%、特异度96.02%，阴性预测值高达99.41%；金胺“O”荧光染色与萋–尼氏抗酸染色特异度均达100.00%，但灵敏度仅为52.38%、42.86%；固体培养法灵敏度57.14%，特异度99.43%。韦恩图分析提示，近半数阳性样本仅可通过DNA检测检出，传统染色法无独立阳性检出病例。结论：结核分枝杆菌DNA检测法综合诊断效能最优，可有效检出传统方法漏检病例。金胺“O”荧光染色、抗酸染色阳性结果确诊可靠性高，但易漏诊，可用于快速初筛。临床可根据检测特点联合多种方法开展诊断，提升结核病诊断准确率。

Abstract: Objective: This paper aims to compare the diagnostic performance of Ziehl-Neelsen acid-fast staining, Auramine “O” fluorescence staining, M. tuberculosis DNA detection, and solid culture for Mycobacterium tuberculosis, and to provide evidence for early tuberculosis diagnosis. Methods: 218 sputum or bronchoalveolar lavage fluid specimens were tested with all four assays. Positive detection rates were recorded; sensitivity, specificity, positive and negative predictive values were compared. Venn diagrams were used to analyze the distribution of positive samples. Results: The positive rate of M. tuberculosis DNA detection was 22.02%, significantly higher than the three traditional methods (P < 0.01). Using clinical diagnosis as the reference, the DNA assay showed a sensitivity of 97.62%, specificity of 96.02%, and a high negative predictive value of 99.41%. Auramine “O” fluorescence staining and Ziehl-Neelsen acid-fast staining both had 100% specificity but low sensitivities (52.38% and 42.86%, respectively). Solid culture had a sensitivity of 57.14% and specificity of 99.43%. Venn analysis showed nearly half of positive cases were only detected by DNA, with no independent positives from traditional staining. Conclusion: The M. tuberculosis DNA assay had the best overall diagnostic performance, effectively identifying cases missed by conventional methods. Auramine “O” fluorescence staining and acid-fast staining have high positive predictive value but poor sensitivity, suitable for rapid screening. Combining multiple methods based on their strengths can improve tuberculosis diagnostic accuracy.