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肺结核患者环腺苷酸反应元件结合蛋白与Y-干扰素基因启动子区的关系
  • 期刊名称:《结核病与胸部肿瘤》
  • 时间:0
  • 分类:R521.7[医药卫生—临床医学;医药卫生—内科学]
  • 作者机构:[1]北京市结核病胸部肿瘤研究所结核病分子生物学研究室,北京101149
  • 相关基金:国家自然科学基金资助项目(30771916);国家重大专项“结核病诊断分子标识研究”2008ZX10003-005)
中文摘要:

目的探讨胸腔积液PEMC中CD4+T淋巴细胞分泌或产生IFN—γ的检测对结核性胸膜炎的辅助诊断价值。方法分离40例结核性胸腔积液(结核组)和30例恶性胸腔积液患者(疾病对照组)PELVIC,经克隆表达的早期分泌抗原靶6(ESAT-6)和培养滤液蛋白10(CFP-10)融合蛋白(简称“E/C”)刺激后,用ELISpot检测分泌IFN—γ细胞的斑点形成细胞(SFC)数量和流式细胞术(FCM)结合胞内细胞因子染色检测产生IFN-γ细胞比率,并比较这2项指标在结核组和疾病对照组间的差异,同时评价2种方法检测IFN-γ对结核性胸膜炎的辅助诊断效能。结果PEMC经E/C刺激后用ELISpot检测结核组SFC数量为205(125-450)SFC/5×10^4PEMC,疾病对照组为5(2~18)SFC/5×10^4PEMC,两组间的差异有统计学意义(u=20.00,P〈0.01);FCM检测结核组CD4+T淋巴细胞产生1FN—Y细胞比率为3.27%(1.81%-7.34%),疾病对照组为0.12%(0.06%-0.46%),两组问的差异有统计学意义(U=45.00,P〈0.01)。ELISpot检测PEMC经E/C刺激后分泌IFN-γ的敏感度为92.5%(37/40)、特异度为80.0%(24/30)、阳性预测值为0.86、阴性预测值为0.89、阳性似然比为4.63、阴性似然比为0.09和准确度为87.1%;FCM检测经E/C刺激后产生IFN-γ的敏感度为87.5%(35/40)、特异度为90.0%(27/30)、阳性预测值为0.92、阴性预测值为0.84、阳性似然比为8.75、阴性似然比为o.14和准确度为88.6%。结论经E/C刺激后,用FCM和ELISpot检测CD4+T淋巴细胞产生和分泌IFN—γ细胞的方法对结核性胸膜炎诊断的敏感度和特异度高,且对结核性胸膜炎有辅助诊断价值。

英文摘要:

Objective To explore the value of IFN- γ produced or secreted by CD4+ T Lymphocytes from pleural effusion mononuclear ceils for the diagnosis of tuberculous pleurisy (plTB) . Methods The PEMCs of 40 patients with tuberculous pleural effusion and 30 patients with malignancy pleural effusion were selected as the tuberculosis and disease control groups, then co-cultured with the early secretory antigenic target 6(ESAT-6) and culture filtered protein 10 (CFP-10) fusion protein (E/C). The numbers of spot forming ceils (SFC) secreting IFN-γ were enumerated by ELISpot and the ratios of cells producing IFN-γ were detected by flow cytometry and intracellular cytokine staining. Moreover, the two indicators were compared between tuberculosis and disease control groups to evaluate the 2 methods detecting IFN-γ in the diagnosis of plTB. Results After E/C stimulation, the numbers of SFC were 205 (125-450) SFC/5 × 10^4 PEMC in tuberculosis group and 5 (2-18) SFC/5 × 10^4 PEMC in disease control group by ELISpot. The difference between two groups was statistically significant (U = 20.00, P 〈0.01). The proportion of IFN- γ -secreting CD4+ T lymphocytes was 3.27% (1.81% - 7.34%) in tuberculosis group and 0.12% (0.06% - 0.46%) in control group detected by FCM. The difference between the two groups was statistically significant (U = 45.00, P 〈0.01). The indicators of ELISpot in detection of IFN-γ which was secreted by PEMC after co-cultured with E/C were as follows: sensitivity 92.5% (37/40), specificity 80.0% (24/30), positive predictive value 0.86, negative predictive value 0.89, positive likelihood ratio 4.63, negative likelihood ratio 0.09 and accuracy 87.1%; and for FCM, they were 87.5% (35/40), 90.0% (27/30), 0.92, 0.84, 8.75 and 0.14, respectively and accuracy 88.6%. Coneluslon After E/C stimulation, the assay for IFN- γ -secreting CD4+ T Lymphocytes by FCM and ELISpot is highly sensitive and specific for diagnosis of plTB as an auxiliary method.

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