University Clinical Research Center-SEREFO Laboratory, University of Sciences, Techniques and Technologies of Bamako, Bamako, Mali.
University Clinical Research Center-SEREFO Laboratory, University of Sciences, Techniques and Technologies of Bamako, Bamako, Mali; Medical Biochemistry, School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Durban, South Africa.
Int J Mycobacteriol. 2023 Apr-Jun;12(2):144-150. doi: 10.4103/ijmy.ijmy_13_23.
Despite recent advances in the development of more sensitive technologies for the diagnosis of tuberculosis (TB), in resource-limited settings, the diagnosis continues to rely on sputum smear microscopy. This is because smear microscopy is simple, cost-efficient and the most accessible tool for the diagnosis of TB. Our study evaluated the performance of light-emitting diode fluorescence microscopy (LED-FM) using auramine/rhodamine (auramine) and the fluorescein di-acetate (FDA) vital stain in the diagnostic of pulmonary TB in Bamako, Mali.
Sputum smear microscopy was conducted using the FDA and auramine/rhodamine staining procedures on fresh samples using LED-FM to evaluate the Mycobacterium TB (MTB) metabolic activity and to predict contagiousness. Mycobacterial culture assay was utilized as a gold standard method.
Out of 1401 TB suspected patients, 1354 (96.65%) were retrieved from database, which were MTB complex culture positive, and 47 (3.40%) were culture negative (no mycobacterial growth observed). Out of the 1354 included patients, 1343 (95.86%), were acid-fast bacillus (AFB) positive after direct FDA staining, 1352 (96.50%) AFB positive after direct Auramine, and 1354 (96.65%) AFB positive with indirect auramine after digestion and centrifugation. Overall, the FDA staining method has a sensitivity of 98.82%, while the sensitivity of Auramine with direct observation was 99.48%, and 99.56% with the indirect examination.
This study showed that, using fresh sputum both auramine/rhodamine and FDA are highly sensitive methods in diagnosing pulmonary TB and could be easily used in countries with limited resource settings.
尽管最近在开发更敏感的结核病（TB）诊断技术方面取得了进展，但在资源有限的环境中，诊断仍然依赖于痰涂片显微镜检查。这是因为涂片显微镜检查简单、具有成本效益，并且是诊断结核病最容易获得的工具。我们的研究评估了在马里巴马科使用发光二极管荧光显微镜（LED-FM）使用吖啶橙/罗丹明（吖啶橙）和荧光素二乙酸酯（FDA）活力染色诊断肺结核的性能。
使用 FDA 和吖啶橙/罗丹明染色程序对新鲜样本进行痰涂片显微镜检查，使用 LED-FM 评估结核分枝杆菌（MTB）代谢活性并预测传染性。分枝杆菌培养检测被用作金标准方法。
从数据库中检索到 1401 例疑似结核病患者，其中 1354 例（96.65%）为 MTB 复合培养阳性，47 例（3.40%）为培养阴性（未观察到分枝杆菌生长）。在包括的 1354 例患者中，直接 FDA 染色后 1343 例（95.86%）为抗酸杆菌（AFB）阳性，直接吖啶橙染色后 1352 例（96.50%）为 AFB 阳性，消化和离心后间接吖啶橙染色后 1354 例（96.65%）为 AFB 阳性。总体而言，FDA 染色方法的敏感性为 98.82%，而直接观察时吖啶橙的敏感性为 99.48%，间接检查时为 99.56%。
本研究表明，使用新鲜痰液，吖啶橙/罗丹明和 FDA 都是诊断肺结核的高度敏感方法，并且可以在资源有限的国家中轻松使用。
