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 当前的SARS-COV-2分子诊断测试主要基于实时定量PCR （qpcr）。 Digital PCR是基于限制稀释的下一代PCR技术 PCR和泊松统计。数字PCR改变了qPCR的指数性，类似物性质 定量成线性的数字信号量化。与QPCR相比，数字PCR提供 几个重要优势，包括其提供绝对量化的能力，容忍度 抑制剂/污染物及其高灵敏度。 最近的两项研究表明，数字PCR提高了提高covid-19的灵敏度。 QPCR错过了。另外，对于某些患者，qPCR的SARS-COV-2感染结果 每天变化。但是与QPCR不同，数字PCR显示出一致的可重复结果。 总体而言，数字PCR被证明可以降低显着错误的阴性结果，即 对于诊断早期或无症状感染或测试康复特别有用 出院前的患者。 QPCR中使用的当前荧光探针，例如FAM（荧光素染料），没有 亮度将使用手机型相机可视化。解决这个问题并启用 数字PCR的简单单元手机读数导致护理点设置，该项目的目的是 将pdots开发为可以提供敏感检测的数字PCR测定的Ultrabright探针 SARS-COV-2。