脑脊液单核细胞ESAT-6、LAM检测和16s RNA基因检测对结核性脑膜炎的诊断价值研究

目的:探讨针对检测脑脊液单核细胞中结核杆菌分泌性的ESAT-6、LAM(lipoarabinomannan)蛋白以及16sRNA基因水平对结核性脑膜炎的诊断价值。方法:在XX医院收集病例45人,实验共分成三个组,分别是结核性脑膜炎病人20人作为结核性脑膜炎组,病毒性脑膜炎5人作为化脓性脑膜炎组,正常没有疾病的人20人作为正常组。分别分离出脑脊液单核细胞,用免疫荧光的方法检测单核细胞内ESAT-6和LAM的蛋白表达水平;另外用实时定量PCR的方法检测结核杆菌16s RNA基因。结果:单核细胞中ESAT-6蛋白在结核性脑膜炎组中有16例阳性,在5例病毒性脑膜炎病人中全阴性;LAM蛋白在结核性脑膜炎组中有14人反应阳性,在病毒性脑膜炎组中有一人阳性,约登指数0.5;定量PCR检测显示,结核性脑膜炎组中16s RNA阳性18人,正常组和病毒性脑膜炎组中都没有检测到,其敏感性为90%,特异性100%,约登指数0.90。结论:测定脑脊液单核细胞中结核杆菌分泌性的ESAT-6、LAM蛋白及16s RNA基因水平对结核性脑膜炎具有重要诊断价值。

Objective: To explore the diagnose value on tuberculous meningitis by analysis the ESAT-6 protein, LAM glycolipid and 16 S RNA gene. Methods: 45 people were recruited in XX hospital. According to the clinical diagnose, we divided into three groups. The first group is 20 cases of tuberculous meningitis, the second group is 5 cases of viral meningitis, and 20 normal people without any kinds of meningitis or neuro diseases as normal control. Collection of the cerebrospinal fluid, and then analysis of ESAT-6 content by immunofluorescence, analysis of LAM content by enzyme-linked immunosorbent assay, and analysis of 16 S RNA gene by Real-Time PCR. Results: Detection of ESAT-6 protein in cerebrospinal fluid monocytoid cells showed that 16/20 cases are positive in tuberculous meningitis group. While, in viral meningitis and normal groups are undetectable. The sensitivity was 80%, the specificity was 100%, and Youden index is 0.8. Further, 14/20 tuberculous meningitis cases were positive of LAM in the cerebrospinal fluid; one viral meningitis case was positive; the normal group was LAM negative. The sensitivity was 70%, the specificity was 80%, and Youden index is 0.5.Moreover, Real-time PCR showed that 18/20 tuberculous meningitis cases were positive of 16 S RNA gene; the rest two groups are undetectable. So the sensitivity was 90%, the specificity was 100%, and Youden index is 0.9. Conclusions: The determination level of ESAT-6, LAM protein and 16 S RNA gene secreted by Mycobacterium tuberculosis in CSF monocytes has important diagnostic value for tuberculous meningitis..