Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence.
Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence.
复制标题
使用报告酶荧光快速诊断结核病。
DOI：
10.1128/jcm.01462-19
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发表时间：
2019
影响因子：
9.4
通讯作者：
Cirillo,JeffreyD
中科院分区：
文献类型：
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作者：
Sule,Preeti;Tilvawala,Ronak;Mustapha,Toriq;Hassounah,Hany;Noormohamed,Aneesa;Kundu,Suprateek;Graviss,EdwardA;Walkup,GrantK;Kong,Ying;Cirillo,JeffreyD
结核病是单一传染源导致人类死亡的最常见原因。由于在早期感染期间痰中存在的细菌数量较少，通常在症状出现后>3至4个月才能诊断。我们创造了一种新的更灵敏的诊断方法，可以在10分钟内进行，无需处理或技术专业知识。该检测试剂盒利用报告酶荧光（REF）中的结核分枝杆菌特异性生物标志物BlaC，该酶已针对临床样本进行了优化，指定为REFtb，沿着更特异的荧光底物CDG-3。我们报告的第一个评价临床标本与结核病诊断，培养和涂片镜检的金标准相比，REFtb检测。REFtb检测允许诊断来自16个不同国家的160名患者，对涂片阳性、培养阳性样本的灵敏度为89%，对涂片阴性、培养阳性样本的灵敏度为88%，特异性为82%。REFtb对结核感染的阴性预测值为93%，阳性预测值为79%。总的来说，这些数据表明需要由第三方使用市售REFtb试剂盒进行更大的准确性研究，以确定将REFtb纳入疑似结核病患者的临床工具箱是否会改善病例识别。如果所有诊断实验室都能获得与我们相似的结果，REFtb将允许对超过85%的患者进行适当的治疗，这些患者在使用当前诊断策略初次就诊时会被遗漏，从而降低疾病进一步传播的可能性。
Tuberculosis is the most frequent cause of death in humans from a single infectious agent. Due to low numbers of bacteria present in sputum during early infection, diagnosis does not usually occur until >3 to 4 months after symptoms develop. We created a new more sensitive diagnostic that can be carried out in 10 min with no processing or technical expertise. This assay utilizes the Mycobacterium tuberculosis-specific biomarker BlaC in reporter enzyme fluorescence (REF) that has been optimized for clinical samples, designated REFtb, along with a more specific fluorogenic substrate, CDG-3. We report the first evaluation of clinical specimens with REFtb assays in comparison to the gold standards for tuberculosis diagnosis, culture and smear microscopy. REFtb assays allowed diagnosis of 160 patients from 16 different countries with a sensitivity of 89% for smear-positive, culture-positive samples and 88% for smear-negative, culture-positive samples with a specificity of 82%. The negative predictive value of REFtb for tuberculosis infection is 93%, and the positive predictive value is 79%. Overall, these data point toward the need for larger accuracy studies by third parties using a commercially available REFtb kit to determine whether incorporation of REFtb into the clinical toolbox for suspected tuberculosis patients would improve case identification. If results similar to our own can be obtained by all diagnostic laboratories, REFtb would allow proper treatment of more than 85% of patients that would be missed during their initial visit to a clinic using current diagnostic strategies, reducing the potential for further spread of disease.