Jiao Wei-Wei, Wang Yi, Wang Gui-Rong, Wang Ya-Cui, Xiao Jing, Sun Lin, Li Jie-Qiong, Wen Shu-An, Zhang Ting-Ting, Ma Qi, Huang Hai-Rong, Shen A-Dong
Key Laboratory of Major Diseases in Children, Ministry of Education, National Key Discipline of Pediatrics (Capital Medical University), National Clinical Research Center for Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Disease, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing, China.
National Tuberculosis Clinical Laboratory, Beijing Key Laboratory for Drug Resistance Tuberculosis Research, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing Chest Hospital, Capital Medical University, Beijing, China.
Front Microbiol. 2019 Sep 13;10:2135. doi: 10.3389/fmicb.2019.02135. eCollection 2019.
Tuberculosis is still a major threat to global public health. Here, a novel diagnosis assay, termed as multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor (MCDA-LFB), was developed to simultaneously detect IS and IS of (MTB) in DNA extracted from reference strain H37Rv and clinical samples. The amplification can be finished within 30 min at a fixed temperature (67°C), thus the whole procedure, including rapid template preparation (15 min), isothermal reaction (30 min) and result reporting (2 min), can be completed within 50 min. The limit of detection of multiplex MCDA assay was 10 fg per reaction. By using the multiplex MCDA protocol, cross-reaction with non-mycobacteria and non-tuberculous mycobacteria (NTM) strains was not observed. Among clinically diagnosed TB patients, the sensitivity of liquid culture, Xpert MTB/RIF and multiplex MCDA assay was 42.0% (50/119), 49.6% (59/119), and 88.2% (105/119), respectively. Among culture positive samples, the sensitivity of Xpert MTB/RIF and multiplex MCDA assay was 86.0% (43/50) and 98.0% (49/50), respectively. Among culture negative samples, the sensitivity of Xpert MTB/RIF and multiplex MCDA assay was 23.2% (16/69) and 81.2% (56/69), respectively. The specificity was 100% (60/60) for Xpert MTB/RIF and 98.3% (59/60) for multiplex MCDA. Therefore, the multiplex MCDA assay for MTB detection is rapid, sensitive and easy to use and may be a promising test for early diagnosis of TB.
结核病仍然是全球公共卫生的重大威胁。在此，开发了一种新型诊断检测方法，称为多重交叉置换扩增结合基于纳米颗粒的侧向流动生物传感器（MCDA-LFB），用于同时检测从参考菌株H37Rv和临床样本中提取的DNA中的结核分枝杆菌（MTB）的IS和IS。扩增可在固定温度（67°C）下30分钟内完成，因此整个过程，包括快速模板制备（15分钟）、等温反应（30分钟）和结果报告（2分钟），可在50分钟内完成。多重MCDA检测的检测限为每个反应10 fg。通过使用多重MCDA方案，未观察到与非结核分枝杆菌和非结核分枝杆菌（NTM）菌株的交叉反应。在临床诊断的结核病患者中，液体培养、Xpert MTB/RIF和多重MCDA检测的敏感性分别为42.0%（50/119）、49.6%（59/119）和88.2%（105/119）。在培养阳性样本中，Xpert MTB/RIF和多重MCDA检测的敏感性分别为86.0%（43/50）和98.0%（49/50）。在培养阴性样本中，Xpert MTB/RIF和多重MCDA检测的敏感性分别为23.2%（16/69）和81.2%（56/69）。Xpert MTB/RIF的特异性为100%（60/60），多重MCDA的特异性为98.3%（59/60）。因此，用于MTB检测的多重MCDA检测快速、灵敏且易于使用，可能是结核病早期诊断的一种有前途的检测方法。
