biostudies-literatureLuo DChinese Academy of Sciences445-453https://www.ebi.ac.uk/biostudies/studies/S-EPMC10173069biostudies-literatureUnknown32High purity is essential in mRNA-based therapeutic applications. A major contaminant of <i>in vitro</i>-transcribed (IVT) mRNA manufacturing is double-stranded RNA (dsRNA), which can induce severe anti-viral immune responses. Detection methods, such as agarose gel electrophoresis, ELISA, and dot-blot assay, are used to detect the existence of dsRNA in IVT mRNA products. However, these methods are either not sensitive enough or time-consuming. To overcome these challenges, we develop a rapid, sensitive, and easy-to-implement colloidal gold nanoparticle-based lateral flow strip assay (LFSA) with sandwich format for the detection of dsRNA from IVT process. dsRNA contaminant can be determined visually on the test strip or quantitatively with a portable optical detector. This method allows for Molecular therapy. Nucleic acidsLateral flow immunoassay for rapid and sensitive detection of dsRNA contaminants in <i>in vitro</i>-transcribed mRNA products.PMC10173069YSBR010Shao FWu ZCestellos-Blanco SZhang JWang SWang DCao YXu DLuo DfalseLateral flow immunoassay for rapid and sensitive detection of dsRNA contaminants in <i>in vitro</i>-transcribed mRNA products.High purity is essential in mRNA-based therapeutic applications. A major contaminant of <i>in vitro</i>-transcribed (IVT) mRNA manufacturing is double-stranded RNA (dsRNA), which can induce severe anti-viral immune responses. Detection methods, such as agarose gel electrophoresis, ELISA, and dot-blot assay, are used to detect the existence of dsRNA in IVT mRNA products. However, these methods are either not sensitive enough or time-consuming. To overcome these challenges, we develop a rapid, sensitive, and easy-to-implement colloidal gold nanoparticle-based lateral flow strip assay (LFSA) with sandwich format for the detection of dsRNA from IVT process. dsRNA contaminant can be determined visually on the test strip or quantitatively with a portable optical detector. This method allows for 2023-01-01T00:00:00Z2023 Jun2025-04-05T09:12:02.147Z2025-04-05T09:12:02.147ZS-EPMC101730693718145010.1016/j.omtn.2023.04.005