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.

项目摘要 目前SARS-CoV-2的分子诊断方法主要是基于实时荧光定量PCR技术 （qPCR）。数字PCR是基于有限稀释、终点检测和定量分析的下一代PCR技术。 PCR和Poisson统计。数字PCR改变了qPCR的指数模拟性质 量化转换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， 抑制剂/污染物，以及其高灵敏度。 最近的两项研究表明，数字PCR提高了检测COVID-19的灵敏度， qPCR缺失。此外，对于某些患者，qPCR检测SARS-CoV-2感染的结果 每天都有变化。但与qPCR不同，数字PCR显示出一致的可重复结果。 总的来说，数字PCR显示出显着减少假阴性结果，这是 特别适用于诊断早期或无症状感染或用于检测恢复期 患者出院前 目前用于qPCR的荧光探针，如FAM（荧光素染料），不具有荧光素标记。 使用手机类型的摄像头可以可视化亮度。为了解决这个问题， 简单的手机读出数字PCR的结果在一个护理点设置，该项目的目的是 将Pdot开发成用于数字PCR检测的超亮探针， SARS-CoV-2