Division of Biomedical Engineering, School of Engineering, University of Glasgow, Oakfield Avenue, Glasgow, G12 8LT, UK.
Public Health England Malaria Reference Laboratory (PHE MRL), Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, UK.
Angew Chem Int Ed Engl. 2016 Dec 5;55(49):15250-15253. doi: 10.1002/anie.201606060. Epub 2016 Aug 24.
We demonstrate, for the first time, the multiplexed determination of microbial species from whole blood using the paper-folding technique of origami to enable the sequential steps of DNA extraction, loop-mediated isothermal amplification (LAMP), and array-based fluorescence detection. A low-cost handheld flashlight reveals the presence of the final DNA amplicon to the naked eye, providing a "sample-to-answer" diagnosis from a finger-prick volume of human blood, within 45 min, with minimal user intervention. To demonstrate the method, we showed the identification of three species of Plasmodium, analyzing 80 patient samples benchmarked against the gold-standard polymerase chain reaction (PCR) assay in an operator-blinded study. We also show that the test retains its diagnostic accuracy when using stored or fixed reference samples.
我们首次展示了使用折纸技术对全血中的微生物物种进行多重检测，该技术可实现 DNA 提取、环介导等温扩增（LAMP）和基于阵列的荧光检测等连续步骤。低成本的手持手电筒可使最终的 DNA 扩增子在肉眼下可见，从而可在 45 分钟内从手指穿刺的人类血液样本中实现“样本到答案”的诊断，用户干预极少。为了演示该方法，我们通过盲法操作员研究，以金标准聚合酶链反应（PCR）检测方法为基准，对 80 个患者样本进行鉴定，证实了该方法能够鉴定三种疟原虫。我们还表明，该测试在使用存储或固定的参考样本时仍然具有诊断准确性。
