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Novel approaches in diagnosing tuberculosis

机译:结核病诊断的新方法

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The WHO declared tuberculosis (TB) a global emergency. An estimated 8-9 million new cases occur each year with 2-3 million deaths. Currently, TB is diagnosed mostly by chest-X ray and staining of the mycobacteria in sputum with a detection limit of 1x104 bacteria /ml. There is an urgent need for better diagnostic tools for TB especially for developing countries. We have validated the electronic nose from TD Technology for the detection of Mycobacterium tuberculosis by headspace analysis of 284 sputum samples from TB patients. We used linear discriminant function analysis resulting in a sensitivity of 75% a specificity of 67% and an accuracy of 69%. Further research is still required to improve the results by choosing more selective sensors and sampling techniques. We used a fast gas chromatography- mass spectrometry method (GC-MS). The automated procedure is based on the injection of sputum samples which are methylated inside the GC injector using thermally assisted hydrolysis and methylation (THM-GC-MS). Hexacosanoic acid in combination with tuberculostearic acid was found to be specific for the presence of M. tuberculosis. The detection limit was similar to microscopy. We found no false positives, all microscopy and culture positive samples were also found positive with the THM-GC-MS method. The detection of ribosomal RNA from the infecting organism offers great potential since rRNA molecules outnumber chromosomal DNA by a factor 1000. It thus may possible to detect the organism without amplification of the nucleic acids (NA). We used a capture and a tagged detector probe for the direct detection of M. tuberculosis in sputum. So far the detection limit is 1x106 bacteria / ml. Currently we are testing a Lab-On-A-Chip Interferometer detection system
机译:谁宣布结核病(TB)全球紧急情况。估计每年均为8-900万新案件,每年发生2-300万人死亡。目前,TB主要被胸X射线诊断,并在痰中染色分枝杆菌,检测限为1×104细菌/ ml。迫切需要更好地诊断TB,特别是对于发展中国家。通过来自TB患者的284个痰样品的前空分析,我们从TD技术验证了来自TD技术的电子鼻子。我们使用了线性判别功能分析,其灵敏度为75%的特异性为67%,精度为69%。仍然需要进一步的研究来通过选择更多选择性传感器和采样技术来改进结果。我们使用了快速气相色谱 - 质谱法(GC-MS)。自动化程序基于注射使用热辅助水解和甲基化(THM-GC-MS)内的GC喷射器内甲基化的痰液样品。六烷烷酸与结核病酸结合的是特异于结核病的存在。检测限类似于显微镜。我们发现没有假阳性,也发现所有显微镜和培养阳性样品也与THM-GC-MS法呈正阳性。来自感染物体的核糖体RNA的检测提供了很大的潜力,因为RRNA分子超过染色体DNA,因此可以在不扩增核酸(NA)的情况下检测生物体。我们使用捕获和标记的探测器探针,用于直接检测痰中的肺结核。到目前为止,检测限是1x106细菌/ ml。目前我们正在测试一个芯片实验室干涉仪检测系统

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