School of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea.
Biosens Bioelectron. 2020 Dec 15;170:112656. doi: 10.1016/j.bios.2020.112656. Epub 2020 Sep 26.
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).
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)。