Evaluation of Aptamer Fluorescence Microscopy in the Diagnosis of Pulmonary Tuberculosis.
Evaluation of Aptamer Fluorescence Microscopy in the Diagnosis of Pulmonary Tuberculosis.
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DOI：
10.1128/spectrum.02602-21
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发表时间：
2022-08-31
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
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痰涂片镜检诊断结核病经受住了时间的考验。但由于其敏感性较低，对临床标本中结核病的阳性检出率不高。为了提高结核分枝杆菌（MTB）检测的显微镜观察灵敏度，我们开发了MTB特异性适配子MA 1。为了进一步提高MA 1与MTB的结合反应性，我们构建了一种新的具有口袋-茎-环结构的衍生适体MA 1 -39，并鉴定其与MTB参考菌株具有高结合反应性。我们建立了一种基于MA 1 -39的MTB适配体荧光显微镜检测方法，并评价了其诊断肺结核的可行性。在56株结核分枝杆菌中，MA 1 -39能有效区分12株非结核分枝杆菌（NTM）参考株、6株NTM分离株和7株其他细菌。接下来，将这种方法应用于来自疑似结核病患者和非结核病对照的169份临床痰液样本。分子检测与临床和细菌学鉴定一起被用作评估适体检测功效的方案。与传统的抗酸染色光镜相比，适体荧光显微镜对临床标本中结核分枝杆菌的检出率更高（48.8%比32.6%），特异性几乎无显著性差异（90.4%比94%）。此外，适体荧光显微镜显示与染色相同的阳性预测值（PPV）（84%对84.9%），但更高的阴性预测值（NPV; 63%对57.3%）。因此，新建立的适体荧光显微镜技术有可能成为结核病微生物诊断的一种可行方法。重要信息我们建立了一种适体荧光显微镜方法来快速检测临床痰液样本中的MTB。适体探针的使用被证明可以显著提高痰涂片镜检的灵敏度。在资源有限的国家，显微镜检查目前是许多实验室的一种快速、简单和非常常用的检测方法，在可预见的将来，它仍将是结核病微生物诊断的主要手段。改进检测技术可以进一步提高这一古老诊断方法的临床应用价值。由于适体荧光显微镜可以提供快速和灵敏的结果，它可能是一个可行的和有用的方法，在资源有限的设置。
Sputum smear microscopy for tuberculosis diagnosis has stood the test of time. However, due to its low sensitivity, the positive detection rate for tuberculosis in clinical specimens is not high. To improve the sensitivity of microscopic observation in Mycobacterium tuberculosis (MTB) detection, we developed the MTB-specific aptamer MA1. To further improve the binding reactivity of MA1 with MTB, we constructed a new derivative aptamer with a pocket-stem-loop-structure, MA1-39, and identified it to have high binding reactivity with the MTB reference strain. We developed an aptamer fluorescence microscopy test for MTB based on MA1-39 and evaluated its feasibility for diagnosing pulmonary tuberculosis. Among 56 tested strains, MA1-39 was proven to effectively discriminate MTB from the control strains, including 12 non-tuberculosis mycobacterial (NTM) reference strains, 6 NTM isolates, and 7 other bacteria. Next, this approach was applied to 169 clinical sputum samples from suspected tuberculosis patients and non-tuberculosis controls. Molecular tests together with both clinical and bacteriological identification were used as a protocol to evaluate the efficacy of aptamer detection. Compared with the traditional acid-fast staining light microscope, the aptamer fluorescence microscope showed a higher detection rate for MTB in clinical specimens (48.8% versus 32.6%), and the specificities of the two techniques had almost no significant difference (90.4% versus 94%). In addition, aptamer fluorescence microscopy showed the same positive predictive value (PPV) as staining (84% versus 84.9%), but a higher negative predictive value (NPV; 63% versus 57.3%). In conclusion, the newly established aptamer fluorescence microscopy approach is likely to be a feasible method for microbiological diagnosis of tuberculosis. IMPORTANCE We established an aptamer fluorescence microscopy approach for rapid detection of MTB in clinical sputum samples. The use of aptamer probes was proven to significantly increase the sensitivity of sputum smear microscopy. In resource-limited countries, microscopy is currently a fast, simple, and very common test method in many laboratories, and it will remain the primary means of microbiological diagnosis of tuberculosis in the foreseeable future. Improving detection techniques can further enhance the clinical application value of this ancient diagnostic method. Since aptamer fluorescence microscopy can provide rapid and sensitive results, it may be a feasible and useful method in resource-limited settings.
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DOI：
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发表时间：
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期刊：
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影响因子：
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DOI：
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发表时间：
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期刊：
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影响因子：
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作者：
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通讯作者：
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