脑脊液改良抗酸染色联合单核细胞内早期分泌抗原靶-6测定用于结核性脑膜炎早期诊断的研究

A study of cerebrospinal fluid modified acid - fast staining combined with early secretary antigentic target 6 for early diagnosis of tuberculous meningitis

目的探寻结核性脑膜炎（TBM）早期诊断的新方法。方法选取徐州医科大学附属医院神经内科2013年5月-2015年2月收治的TBM患者38例及其他中枢神经系统感染患者38例。所有病例入院后24-48h内，腰椎穿刺脑脊液（CSF）并予常规检测，同时采用传统集菌法涂片抗酸染色检测结核分枝杆菌（MTB）、改良抗酸染色检测MTB、免疫细胞化学染色及酶联免疫吸附法（ELISA）测定早期分泌抗原靶-6（ESAT-6）。结果①改良抗酸染色检测MTB的灵敏度（Se）达71．1％，与传统抗酸染色（Se为2．6％）比较，差异有统计学意义（P=0．000）；改良抗酸染色图片中，单核细胞、中性粒细胞内见到MTB，同时淋巴细胞内也发现MTB；在同一样本中，MTB以短棒状、点状、球形及细长弯曲型等多种形态存在。②免疫细胞化学染色诊断TBM的Se为84．2％，Sp为89．5％；单核细胞表达ESAT-6最为常见，中性粒细胞、淋巴细胞内也存在ESAT-6表达现象。③CSF改良抗酸染色联合免疫细胞化学ESAT-6检测诊断TBM的Se为89．5％，与改良抗酸染色比较，差异有统计学意义（P=0．044）；联合检测的Sp为89．5％，与免疫细胞化学染色比较差异无统计学意义（P=0．169）。结论CSF改良抗酸染色联合单核细胞内ESAT-6测定可以提高诊断TBM的灵敏度。

Objective To explore new laboratory methods for early diagnosis of tuberculous meningitis （TBM）. Methods A total of 38 tuberculous meningitis patients and another 38 patients with other central nervous system infec- tions who were admitted into our hospital from May 2013 to February 2015 were included. After admission for 24 to 48 h, all patients underwent lumbar puncture to collect cerebrospinal fluid （CSF） for routine examination. Meanwhile, Myco- bacterium tuberculosis （MTB） was detected by the traditional acid - fast staining method and a modified acid - fast stai- ning. The levels of early secretary antigentic target 6 （ ESAT 6） were determined by ELISA. Results The sensitivity of the modified acid- fast staining was 71.1%, which was remarkably different from that of the traditional method （P = 0.000）. MTB were found in monocytes, neutrophils and lymphocytes, according to the modified acid - fast staining ima- ges. MTB generally exist in short rod, point -like, spherical and elongated curved shapes. The sensitivity and specificity of immunocytochemistry staining for TBM diagnosis was 84.2% and 89.5% , respectively. ESAT - 6 was generally ex- pressed in monocytes, while its presence could be found in both neutrophils and lymphocytes. The sensitivity of a com- bined method of CSF modified acid - fast staining and ESAT - 6 for TBM diagnosis was 89.5%, which was statistically different from that using the modified acid - fast staining alone （ P = 0. 044）. The specificity of the combined method was 89.5%, which was not statistically different from that using immunocytochemistry staining （P = 0. 169）. Conclusions The combination of CSF modified acid - fast staining and ESAT 6 within monocytes can improve the sensitivity for earlydiagnosis of TBM.