Pdot-Enabled Point-of-Care Digital PCR for Sensitive Detection of SARS-CoV-2

支持 Pdot 的床旁数字 PCR 用于 SARS-CoV-2 的灵敏检测

• 批准号: 10165078
• 负责人: Jiangbo Yu
• 金额: $ 41.02万
• 依托单位: LAMPROGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10165078
• 关键词: 2019-nCoV; Address; Biological Assay; COVID-19; Cellular Phone; Detection; Dyes; Early Diagnosis; Fluorescein; Fluorescent Probes; Infection; Methods; Molecular Diagnosis; Nature; Patients; Reproducibility of Results; Signal Transduction; Technology; Testing; Time; analog; base; digital; improved; inhibitor/antagonist; next generation; point of care; statistics

Project Summary Current SARS-CoV-2 molecular diagnosis tests mostly are based on real-time quantitative PCR (qPCR). Digital PCR is a next-generation PCR technology based on limiting dilution, end-point PCR, and Poisson statistics. Digital PCR transforms the exponential, analog nature of qPCR quantification into a linear, digital signal quantification. Compared to qPCR, digital PCR offers several important advantages, including its ability to provide absolute quantification, tolerance to inhibitors/contaminants, and its high sensitivity. Two recent studies showed digital PCR has improved sensitivity for picking up COVID-19 that qPCR had missed. In addition, for some patients, results on SARS-CoV-2 infections from qPCR varied from day to day. But unlike qPCR, digital PCR showed consistent reproducible results. Overall, digital PCR was shown to reduce significantly false-negative results, which is particularly useful for diagnosing early or asymptomatic infections or for testing convalescent patients before discharge. Current fluorescent probes used in qPCR, such as FAM (a fluorescein dye), do not have the brightness to be visualized with a cell-phone type camera. To address this issue and to enable simple cell-phone readout of digital PCR results in a point-of-care setting, this project aims to develop Pdots into ultrabright probes for digital PCR assays that can provide sensitive detection of SARS-CoV-2.

项目概要 目前的SARS-CoV-2分子诊断检测大多基于实时定量PCR （qPCR）。数字PCR是基于有限稀释、终点检测的下一代PCR技术 PCR 和泊松统计。数字 PCR 改变了 qPCR 的指数、模拟性质 量化为线性数字信号量化。与 qPCR 相比，数字 PCR 提供 几个重要的优点，包括其提供绝对定量的能力、对 抑制剂/污染物及其高灵敏度。 最近的两项研究表明，数字 PCR 提高了检测 COVID-19 的灵敏度， qPCR 已错过。此外，对于某些患者，qPCR 检测 SARS-CoV-2 感染的结果 每天都有所不同。但与 qPCR 不同的是，数字 PCR 显示出一致的可重复结果。 总体而言，数字 PCR 可以显着减少假阴性结果，即 对于诊断早期或无症状感染或测试恢复期特别有用 患者出院前。 目前 qPCR 中使用的荧光探针，例如 FAM（一种荧光素染料），不具备 使用手机类型的相机可视化亮度。为了解决这个问题并启用 在护理点环境中通过手机简单读出数字 PCR 结果，该项目旨在 将 Pdot 开发成用于数字 PCR 测定的超亮探针，可提供灵敏的检测 SARS-CoV-2。