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Here we present the optimization of highly cost-effective in-house produced enzymes, and we benchmark their performance against commercial alternatives. We explore the compatibility between multiple DNA polymerases with high strand-displacement activity and thermostable reverse transcriptases required for RT-LAMP. We optimize reaction conditions and demonstrate their applicability using both synthetic RNA and clinical patient samples. Finally, we validate the optimized RT-LAMP assay for the detection of SARS-CoV-2 in unextracted heat-inactivated nasopharyngeal samples from 184 patients. 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X. Yin is the co-founder of Biotech & Biomedicine Science (Shenyang) Co. Ltd. S. Ye and X. Liu are employees of Biotech & Biomedicine Science (Shenyang) Co. Ltd. All other authors declare no potential conflict of interest..","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"1820"}}