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追求灵敏度:从 Raindance ddPCR 平台上的病毒核酸检测和定量中获得的经验教训。

In pursuit of sensitivity: Lessons learned from viral nucleic acid detection and quantification on the Raindance ddPCR platform.

机构信息

AIDS and Cancer Virus Program, Frederick National Laboratory for Cancer Research, Frederick, MD 21702, United States.

出版信息

Methods. 2022 May;201:82-95. doi: 10.1016/j.ymeth.2021.04.008. Epub 2021 Apr 9.

Abstract

Sensitive PCR detection of viral nucleic acids plays a critical role in infectious disease research, diagnosis and monitoring. In the context of SARS-CoV-2 detection, recent reports indicate that digital PCR-based tests are significantly more sensitive than traditional qPCR tests. Numerous factors can influence digital PCR reaction sensitivity. In this review, using a model for human HIV infection and the Raindance ddPCR platform as an example, we describe technical aspects that contribute to sensitive viral signal detection in DNA and RNA from tissue samples, which often harbor viral reservoirs and serve as better predictors of disease outcome and indicators of treatment efficacy.

摘要

灵敏的 PCR 检测在传染病研究、诊断和监测中发挥着关键作用。在 SARS-CoV-2 检测方面,最近的报告表明基于数字 PCR 的检测方法比传统 qPCR 检测方法更灵敏。许多因素会影响数字 PCR 反应的灵敏度。在本综述中,我们以人类 HIV 感染模型和 Raindance ddPCR 平台为例,描述了有助于从组织样本中灵敏检测 DNA 和 RNA 中病毒信号的技术方面,这些样本通常含有病毒储存库,并且是更好的疾病结局预测因子和治疗效果指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20de/8526132/241b9d6f10a6/gr1_lrg.jpg

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