应用荧光显微镜检测痰液中的抗酸杆菌作为资源有限国家诊断肺结核的一种有能力的替代工具。

Use of light-emitting diode fluorescence microscopy to detect acid-fast bacilli in sputum as proficient alternative tool in the diagnosis of pulmonary tuberculosis in countries with limited resource settings.

机构信息

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.

出版信息

Int J Mycobacteriol. 2023 Apr-Jun;12(2):144-150. doi: 10.4103/ijmy.ijmy_13_23.

DOI:10.4103/ijmy.ijmy_13_23

PMID:37338475

Abstract

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

背景

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。

方法

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。

结果

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%。