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In this study, we developed a single-molecule desktop sequencer, GenoCare 1600 (GenoCare), which utilizes amplification-free library preparation and two-color sequencing-by-synthesis chemistry, making it more user-friendly compared with previous single-molecule sequencing platforms for clinical use. Using the GenoCare platform, we sequenced an Escherichia coli standard sample and achieved a consensus accuracy exceeding 99.99%. We also evaluated the sequencing performance of this platform in microbial mixtures and coronavirus disease 2019 (COVID-19) samples from throat swabs. Our findings indicate that the GenoCare platform allows for microbial quantitation, sensitive identification of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, and accurate detection of virus mutations, as confirmed by Sanger sequencing, demonstrating its remarkable potential in clinical application.<\/jats:p>","DOI":"10.1093\/gpbjnl\/qzae006","type":"journal-article","created":{"date-parts":[[2024,1,12]],"date-time":"2024-01-12T06:06:19Z","timestamp":1705039579000},"source":"Crossref","is-referenced-by-count":1,"title":["A Two-color Single-molecule Sequencing Platform and Its Clinical Applications"],"prefix":"10.1093","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0028-8967","authenticated-orcid":false,"given":"Fang","family":"Chen","sequence":"first","affiliation":[{"name":"GeneMind Biosciences Co., Ltd. , Shenzhen 518000, 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