Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional tools. During the 2020 global health crisis, efforts intensified to optimize the production and delivery of molecular diagnostic kits for detecting SARS-CoV-2. During this period, RT-LAMP emerged as a significant focus. However, the thermolability of the reagents used in this technique necessitates special low-temperature infrastructure for transport, storage, and conservation. These requirements limit distribution capacity and necessitate cost-increasing adaptations. Consequently, this report details the development of a lyophilization protocol for reagents in a colorimetric RT-LAMP diagnostic kit to detect SARS-CoV-2, facilitating room-temperature transport and storage. We conducted tests to identify the ideal excipients that maintain the molecular integrity of the reagents and ensure their stability during room-temperature storage and transport. The optimal condition identified involved adding 5% PEG 8000 and 75 mM trehalose to the RT-LAMP reaction, which enabled stability at room temperature for up to 28 days and yielded an analytical and diagnostic sensitivity and specificity of 83.33% and 90%, respectively, for detecting SARS-CoV-2. This study presents the results of a lyophilized colorimetric RT-LAMP COVID-19 detection assay with diagnostic sensitivity and specificity comparable to RT-qPCR, particularly in samples with high viral load.

涉及核酸（DNA 和 RNA）的分子诊断被认为是功能极其强大的工具。 2020 年全球健康危机期间，我们加大力度优化用于检测 SARS-CoV-2 的分子诊断试剂盒的生产和交付。在此期间，RT-LAMP 成为一个重要的焦点。然而，该技术中使用的试剂的热稳定性需要特殊的低温基础设施来运输、储存和保存。这些要求限制了配送能力，并且需要进行成本增加的调整。因此，本报告详细介绍了比色 RT-LAMP 诊断试剂盒中试剂的冻干方案的开发，以检测 SARS-CoV-2，从而促进室温运输和储存。我们进行了测试，以确定能够保持试剂分子完整性并确保其在室温储存和运输过程中稳定性的理想赋形剂。确定的最佳条件是在 RT-LAMP 反应中添加 5% PEG 8000 和 75 mM 海藻糖，可在室温下稳定长达 28 天，分析和诊断灵敏度和特异性分别为 83.33% 和 90%。用于检测 SARS-CoV-2。本研究展示了冻干比色 RT-LAMP COVID-19 检测分析的结果，其诊断灵敏度和特异性与 RT-qPCR 相当，特别是在病毒载量较高的样本中。