通过基于电泳的 RNA 富集技术灵敏检测空气传播病毒的实用工艺

IF 10.61 Q3 Biochemistry, Genetics and Molecular Biology Biosensors and Bioelectronics: X Pub Date : 2024-05-10 DOI:10.1016/j.biosx.2024.100488
Huifeng Du , Simone Bruno , Kalon J. Overholt , Sebastian Palacios , Hsin-Ho Huang , Carlos Barajas , Ben Gross , Cindy Lee , Haley K. Evile , Nuno Rufino de Sousa , Antonio Gigliotti Rothfuchs , Domitilla Del Vecchio
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引用次数: 0

摘要

灵敏的现场病毒检测是及时采取措施防止空气传播病毒的必要条件,因为超低的空气中病毒浓度很容易通过吸入感染个人。在此,我们考虑采用一种可现场使用的生物传感工艺,其中包含一个快速 RNA 富集步骤,以便在 RT-qPCR 之前将病毒 RNA 浓缩到一个小体积中。富集方法在 RT-qPCR 兼容的缓冲液中使用电泳,只需 10 分钟就能将 RNA 浓缩近 5 倍。为了使这一性能符合实际情况,我们分析了一种低成本、可现场使用的生物传感过程的最小检测浓度,该过程使用静电沉淀进行空气采样,加热进行病毒 RNA 提取,然后进行 RNA 富集,最后进行 RT-qPCR。通过富集,我们估计空气中的病毒浓度为 5,654 基因组拷贝 (gc)/m3,检出率为 100%;空气中的病毒浓度为 4,221 gc/m3,检出率为 78.6%。鉴于流感和 SARS-CoV-2 等常见病毒在多个室内空间的浓度介于 5,800 至 37,000 gc/m3 之间,我们得出结论:富集后的检测灵敏度足以满足实际应用的需要。
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A fieldable process for sensitive detection of airborne viruses via electrophoresis-based RNA enrichment

Sensitive on-site virus detection is a requirement for timely action against the spread of airborne infection since ultra-low in-air viral concentrations can readily infect individuals when inhaled. Here, we consider a fieldable biosensing process that incorporates a fast RNA enrichment step in order to concentrate viral RNA in a small volume prior to RT-qPCR. The enrichment approach uses electrophoresis in an RT-qPCR-compatible buffer, and allows for concentration of RNA by nearly 5-fold within only 10 min. In order to place this performance into context, we analyzed the minimum detectable concentration of a low-cost, fieldable, biosensing process that uses electrostatic precipitation for air sampling, heating for viral RNA extraction, and then RNA enrichment, followed by RT-qPCR. With enrichment, we estimated an in-air concentration of 5,654 genome copies (gc)/m3 with a 100% detection rate and an in-air concentration of 4,221 gc/m3 with a 78.6% detection rate. Given that the concentrations of common viruses, such as influenza and SARS-CoV-2, in several indoor spaces are between 5,800 and 37,000 gc/m3, we conclude that enrichment allows a detection that is sufficiently sensitive for practical applications.

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来源期刊
Biosensors and Bioelectronics: X
Biosensors and Bioelectronics: X Biochemistry, Genetics and Molecular Biology-Biophysics
CiteScore
4.60
自引率
0.00%
发文量
166
审稿时长
54 days
期刊介绍: Biosensors and Bioelectronics: X, an open-access companion journal of Biosensors and Bioelectronics, boasts a 2020 Impact Factor of 10.61 (Journal Citation Reports, Clarivate Analytics 2021). Offering authors the opportunity to share their innovative work freely and globally, Biosensors and Bioelectronics: X aims to be a timely and permanent source of information. The journal publishes original research papers, review articles, communications, editorial highlights, perspectives, opinions, and commentaries at the intersection of technological advancements and high-impact applications. Manuscripts submitted to Biosensors and Bioelectronics: X are assessed based on originality and innovation in technology development or applications, aligning with the journal's goal to cater to a broad audience interested in this dynamic field.
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