A novel reverse-transcriptase loop mediated amplification (RT-LAMP) method targeting genes encoding the Spike (S) protein and RNA-dependent RNA polymerase (RdRP) of SARS-CoV-2 has been developed. The LAMP assay achieves a comparable limit of detection (25–50 copies per reaction) to commonly used RT-PCR protocols using clinical samples quantified by digital droplet PCR. Precision, cross-reactivity, inclusivity, and limit of detection studies were performed according to regulatory standards. Clinical validation of dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 98.48 % (95 % CI 91.84%–99.96%) and NPA 100.00 % (95 % CI 93.84%–100.00%) based on the E gene and N2 gene reference RT-PCR methods. The method has implications for development of point of care technology using isothermal amplification.

一种新型逆转录酶环介导扩增 (RT-LAMP) 方法针对编码 SARS-CoV-2 的 Spike (S) 蛋白和 RNA 依赖性 RNA 聚合酶 (RdRP) 的基因。LAMP 检测使用通过数字液滴 PCR 定量的临床样本，实现了与常用 RT-PCR 方案相当的检测限（每个反应 25-50 个拷贝）。精密度、交叉反应性、包容性和检测限研究是根据监管标准进行的。双靶点 RT-LAMP（S 和 RdRP 基因）的临床验证实现了基于 E 基因的 PPA 98.48 % (95 % CI 91.84%–99.96%) 和 NPA 100.00 % (95 % CI 93.84%–100.00%)和N2基因参考RT-PCR方法。该方法对使用等温扩增的护理点技术的发展具有影响。