Abstract:

Objective  To explore the value of detecting early secretory antigenic target-6 (ESAT-6) in cerebrospinal fluid (CSF) monocytes by laser scanning confocal microscopy (LSCM) techniques in early diagnosis of tuberculous meningitis (TBM), and to find a more specific method for early diagnosis.  Methods  Double immunofluorescence staining was applied to detect ESAT-6, and LSCM was used to observe the dyeing result of CSF monocytes and to analyze three-dimensional images. ESAT-6 positive cells will present red fluorescence cytoplasm and blue fluorescence nuclei, and negative cells present blue fluorescence nuclei while the cytoplasm is not stained.  Results  ESAT-6 was mainly expressed with red fluorescence in cytoplasm of CSF monocytes in patients with TBM. Among 35 cases of TBM patients, there were 28 patients (80%) ESAT-6 positive, one case (2.86%) ESAT-6 positive in the control group, and the difference was statistically significant (χ² = 42.918, P = 0.000). ESAT-6 sensitivity was 80% and specificity was 97.14%.  Conclusions  The detection of ESAT-6 can provide a certain basis for the early diagnosis of TBM and has high specificity. LSCM techniques can realize simultaneously the observation of multiple fluorescence and provide clear tomography and three-dimensional images. The cytological research has been improved to a new level.

Key words:

摘要：

目的 探讨激光扫描共聚焦显微镜（LSCM）技术检测脑脊液单核细胞早期分泌抗原靶-6（ESAT-6）在结核性脑膜炎早期诊断中的应用价值，为早期诊断寻找一种更特异性的检测方法。方法 采用免疫荧光双标法检测ESAT-6 抗原，LSCM 观察脑脊液单核细胞着色情况（ESAT-6 阳性细胞胞核呈蓝色荧光、胞质呈红色荧光，ESAT-6 阴性细胞胞核呈蓝色荧光、胞质不着色）并分析三维成像结果。结果 ESAT-6 抗原主要表达于结核性脑膜炎患者脑脊液单核细胞胞质，呈红色荧光。结核性脑膜炎组有28 例（80%）ESAT-6 阳性、对照组仅1 例（2.86%）阳性，组间差异具有统计学意义（χ² = 42.918，P = 0.000）；ESAT-6 检测灵敏度80%，特异度97.14%。结论 ESAT-6 抗原检测可为结核性脑膜炎的早期诊断提供一定依据，且特异性较高。LSCM 技术可实现多重荧光同时观察并形成清晰的断层扫描和三维图像，把细胞学研究提高到新水平。

关键词: 结核, 脑膜, 脑脊髓液, 单核细胞, 荧光抗体技术, 显微镜检查, 共焦