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• W4283278313 endingPage "9379" @default.
• W4283278313 startingPage "9372" @default.
• W4283278313 abstract "Polymerase chain reaction (PCR)-based diagnostic testing is the gold standard method for pathogen identification (ID) with recent developments enabling automated PCR tests for point-of-care (POC) use. However, multiplexed identification of several pathogens in PCR assays typically requires optics for an equivalent number of fluorescence channels, increasing instrumentation’s complexity and cost. In this study, we first developed ratiometric PCR that surpassed one target per color barrier to allow multiplexed identification while minimizing optical components for affordable POC use. We realized it by amplifying pathogenic targets with fluorescently labeled hydrolysis probes with a specific ratio of red-to-green fluorophores for each bacterial species. We then coupled ratiometric PCR and automated magnetic beads-based sample preparation within a thermoplastic cartridge and a portable droplet magnetofluidic platform. We named the integrated workflow POC-ratioPCR. We demonstrated that the POC-ratioPCR could detect one out of six bacterial targets related to urinary tract infections (UTIs) in a single reaction using only two-color channels. We further evaluated POC-ratioPCR using mock bacterial urine samples spiked with good agreement. The POC-ratioPCR presents a simple and effective method for enabling broad-based POC PCR identification of pathogens directly from crude biosamples with low optical instrumentation complexity." @default.
• W4283278313 created "2022-06-23" @default.
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• W4283278313 date "2022-06-22" @default.
• W4283278313 modified "2023-10-15" @default.
• W4283278313 title "Ratiometric PCR in a Portable Sample-to-Result Device for Broad-Based Pathogen Identification" @default.
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• W4283278313 doi "https://doi.org/10.1021/acs.analchem.2c01357" @default.
• W4283278313 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35730588" @default.
• W4283278313 hasPublicationYear "2022" @default.
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