Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Methods Mol Biol. 2024;2822:39-50. doi: 10.1007/978-1-0716-3918-4_4.
RT-LAMP is an effective alternative to RT-PCR-based diagnostics, offering high specificity, sensitivity, and rapid results. One notable advantage is the robustness of its enzymes, allowing for direct amplification from crude samples without the need for prior isolation of RNA. Colorimetric LAMP is particularly attractive as it eliminates the need for complex instrumentation, making it suitable for point-of-care applications. Here, we present a comprehensive step-by-step protocol for establishing an RT-LAMP-based test for direct detection of SARS-CoV-2 genomic RNA in saliva samples using different colorimetric detection methods. Importantly, this versatile test can be easily adapted to detect emerging pathogens.
RT-LAMP 是一种基于 RT-PCR 的诊断方法的有效替代方法,具有高特异性、高灵敏度和快速结果的特点。一个显著的优点是其酶的稳健性,允许直接从粗提样本中进行扩增,而无需事先分离 RNA。比色 LAMP 特别有吸引力,因为它消除了对复杂仪器的需求,使其适用于即时检测应用。在这里,我们提供了一个全面的分步协议,用于建立一种基于 RT-LAMP 的测试方法,用于直接检测唾液样本中的 SARS-CoV-2 基因组 RNA,使用不同的比色检测方法。重要的是,这种多功能的测试可以很容易地适应新兴病原体的检测。