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一种用于快速感测空气中冠状病毒和流感病毒的空气采样和同时浓缩的集成系统。

An integrated system of air sampling and simultaneous enrichment for rapid biosensing of airborne coronavirus and influenza virus.

机构信息

School of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea.

出版信息

Biosens Bioelectron. 2020 Dec 15;170:112656. doi: 10.1016/j.bios.2020.112656. Epub 2020 Sep 26.

Abstract

Point-of-care risk assessment (PCRA) for airborne viruses requires a system that can enrich low-concentration airborne viruses dispersed in field environments into a small volume of liquid. In this study, airborne virus particles were collected to a degree above the limit of detection (LOD) for a real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). This study employed an electrostatic air sampler to capture aerosolized test viruses (human coronavirus 229E (HCoV-229E), influenza A virus subtype H1N1 (A/H1N1), and influenza A virus subtype H3N2 (A/H3N2)) in a continuously flowing liquid (aerosol-to-hydrosol (ATH) enrichment) and a concanavalin A (ConA)-coated magnetic particles (CMPs)-installed fluidic channel for simultaneous hydrosol-to-hydrosol (HTH) enrichment. The air sampler's ATH enrichment capacity (EC) was evaluated using the aerosol counting method. In contrast, the HTH EC for the ATH-collected sample was evaluated using transmission-electron-microscopy (TEM)-based image analysis and real-time qRT-PCR assay. For example, the ATH EC for HCoV-229E was up to 67,000, resulting in a viral concentration of 0.08 PFU/mL (in a liquid sample) for a viral epidemic scenario of 1.2 PFU/m (in air). The real-time qRT-PCR assay result for this liquid sample was "non-detectable" however, subsequent HTH enrichment for 10 min caused the "non-detectable" sample to become "detectable" (cycle threshold (CT) value of 33.8 ± 0.06).

摘要

即时定量逆转录聚合酶链反应(qRT-PCR)检测点风险评估(PCRA)需要一种能够将空气中低浓度病毒浓缩到小体积液体中的系统。本研究采用静电空气采样器,在连续流动的液体(气溶胶到水溶胶(ATH)浓缩)和气溶胶收集物中捕获气溶胶化的测试病毒(人冠状病毒 229E(HCoV-229E)、甲型流感病毒亚型 H1N1(A/H1N1)和甲型流感病毒亚型 H3N2(A/H3N2)),同时在涂有伴刀豆球蛋白 A(ConA)的磁性颗粒(CMP)安装的流道中进行水溶胶到水溶胶(HTH)浓缩。ATH 浓缩效率(EC)是通过气溶胶计数方法评估的。相反,ATH 收集样品的 HTH EC 是通过基于透射电子显微镜(TEM)的图像分析和实时 qRT-PCR 检测来评估的。例如,HCoV-229E 的 ATH EC 高达 67000,导致在病毒流行情景下(空气中 1.2 PFU/m)病毒浓度为 0.08 PFU/mL(在液体样品中)。然而,实时 qRT-PCR 检测该液体样品的结果为“未检出”,随后进行 10 分钟的 HTH 浓缩后,使“未检出”的样品变为“可检出”(循环阈值(CT)值为 33.8±0.06)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6133/7518959/b69b98c2188c/gr1_lrg.jpg

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