Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Jilin, P.R. China.
Angew Chem Int Ed Engl. 2017 Jan 19;56(4):992-996. doi: 10.1002/anie.201609108. Epub 2016 Dec 19.
The detection of nucleic acid biomarkers for point-of-care (POC) diagnostics is currently limited by technical complexity, cost, and time constraints. To overcome these shortcomings, we have combined loop-mediated isothermal amplification (LAMP), programmable toehold-mediated strand-exchange signal transduction, and standard pregnancy test strips. The incorporation of an engineered hCG-SNAP fusion reporter protein (human chorionic gonadotropin-O -alkylguanine-DNA alkyltransferase) led to LAMP-to-hCG signal transduction on low-cost, commercially available pregnancy test strips. Our assay reliably detected as few as 20 copies of Ebola virus templates in both human serum and saliva and could be adapted to distinguish a common melanoma-associated SNP allele (BRAF V600E) from the wild-type sequence. The methods described are completely generalizable to many nucleic acid biomarkers, and could be adapted to provide POC diagnostics for a range of pathogens.
用于即时检测（POC）诊断的核酸生物标志物的检测目前受到技术复杂性、成本和时间限制的限制。为了克服这些缺点，我们将环介导等温扩增（LAMP）、可编程的触发介导的链交换信号转导以及标准妊娠测试条结合在一起。引入工程化的 hCG-SNAP 融合报告蛋白（人绒毛膜促性腺激素-O-烷基鸟嘌呤-DNA 烷基转移酶）可在低成本、市售的妊娠测试条上进行 LAMP 到 hCG 的信号转导。我们的检测方法可靠地检测到人类血清和唾液中低至 20 个拷贝的埃博拉病毒模板，并且可以适应区分常见的黑色素瘤相关 SNP 等位基因（BRAF V600E）与野生型序列。所描述的方法完全适用于许多核酸生物标志物，并且可以适应提供一系列病原体的 POC 诊断。
