ArticleNucleic acids research2023
Accurate quantification of DNA using on-site PCR (osPCR) by characterizing DNA amplification at single-molecule resolution.
Article in Nucleic acids research, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Application-focused validation of a viability-selective PMA-qPCR assay for selected skin-associated and cosmetic-relevant bacteria.Frontiers in microbiology · 2026Article
- Ultrasensitive detection of atrazine by Schottky junction photoelectrochemical aptamer sensor based on signal amplification by cascade catalysis of CRISPR/Cas12a and G-quadruplex/hemin DNAzyme.Mikrochimica acta · 2025Article
- Impact of dental pulp cells-derived small extracellular vesicles on the properties and behavior of dental pulp cells: an in-vitro study.BMC oral health · 2025Article
- Rapid and simple detection ofFrontiers in microbiology · 2025Article
- Advances and challenges of mpox detection technology.Biosafety and health · 2024Review
Corrections and comments
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Authors and funding
16 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Despite the need in various applications, accurate quantification of nucleic acids still remains a challenge. The widely-used qPCR has reduced accuracy at ultralow template concentration and is susceptible to nonspecific amplifications. The more recently developed dPCR is costly and cannot handle high-concentration samples. We combine the strengths of qPCR and dPCR by performing PCR in silicon-based microfluidic chips and demonstrate high quantification accuracy in a large concentration range. Importantly, at low template concentration, we observe on-site PCR (osPCR), where only certain sites of the channel show amplification. The sites have almost identical ct values, showing osPCR is a quasi-single molecule phenomenon. Using osPCR, we can measure both the ct values and the absolute concentration of templates in the same reaction. Additionally, osPCR enables identification of each template molecule, allowing removal of nonspecific amplification during quantification and greatly improving quantification accuracy. We develop sectioning algorithm that improves the signal amplitude and demonstrate improved detection of COVID in patient samples.
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Registered trials
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