Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany.
Department of Laboratory Medicine, Hospital St. Georg, Leipzig, Germany.
Nat Commun. 2021 Mar 5;12(1):1467. doi: 10.1038/s41467-021-21627-0.
Efforts to contain the spread of SARS-CoV-2 have spurred the need for reliable, rapid, and cost-effective diagnostic methods which can be applied to large numbers of people. However, current standard protocols for the detection of viral nucleic acids while sensitive, require a high level of automation and sophisticated laboratory equipment to achieve throughputs that allow whole communities to be tested on a regular basis. Here we present Cap-iLAMP (capture and improved loop-mediated isothermal amplification) which combines a hybridization capture-based RNA extraction of gargle lavage samples with an improved colorimetric RT-LAMP assay and smartphone-based color scoring. Cap-iLAMP is compatible with point-of-care testing and enables the detection of SARS-CoV-2 positive samples in less than one hour. In contrast to direct addition of the sample to improved LAMP (iLAMP), Cap-iLAMP prevents false positives and allows single positive samples to be detected in pools of 25 negative samples, reducing the reagent cost per test to ~1 Euro per individual.
努力控制 SARS-CoV-2 的传播促使人们需要可靠、快速且具有成本效益的诊断方法，以便能够对大量人群进行检测。然而，目前用于检测病毒核酸的标准方法虽然灵敏，但需要高度自动化和复杂的实验室设备才能实现高通量，从而能够定期对整个社区进行检测。在这里，我们提出了 Cap-iLAMP（捕获和改进的环介导等温扩增），它将基于杂交捕获的咽漱洗液 RNA 提取与改进的比色 RT-LAMP 检测和基于智能手机的比色评分相结合。Cap-iLAMP 与即时检测兼容，并能够在不到一个小时的时间内检测到 SARS-CoV-2 阳性样本。与直接将样品加入改进的 LAMP（iLAMP）相比，Cap-iLAMP 可以防止假阳性，并允许在 25 个阴性样本的混合样本中检测到单个阳性样本，从而将每个样本的检测试剂成本降低至约 1 欧元。
