Microfluidic affinity selection of active SARS-CoV-2 virus particles
We report a microfluidic assay to select active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles (VPs), which were defined as intact particles with an accessible angiotensin-converting enzyme 2 receptor binding domain (RBD) on the spike (S) protein, from clinical samples. Affinity selection of SARS-CoV-2 particles was carried out using injection molded microfluidic chips, which allow for high-scale production to accommodate large-scale screening. The microfluidic contained a surface-bound aptamer directed against the virus's S protein RBD to affinity select SARS-CoV-2 VPs. Following selection (~94% recovery), the VPs were released from the chip's surface using a blue light light-emitting diode (89% efficiency). Selected SARS-CoV-2 VP enumeration was carried out using reverse transcription quantitative polymerase chain reaction. The VP selection assay successfully identified healthy donors (clinical specificity = 100%) and 19 of 20 patients with coronavirus disease 2019 (COVID-19) (95% sensitivity). In 15 patients with COVID-19, the presence of active SARS-CoV-2 VPs was found. The chip can be reprogrammed for any VP or exosomes by simply changing the affinity agent.
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Gamage, S. S., Pahattuge, T. N., Wijerathne, H., Childers, K., Vaidyanathan, S., Athapattu, U. S., Zhang, L., Zhao, Z., Hupert, M. L., Muller, R. M., Muller-Cohn, J., Dickerson, J., Dufek, D., Geisbrecht, B. V., Pathak, H., Pessetto, Z., Gan, G. N., Choi, J., Park, S., Godwin, A. K., Witek, M. A., & Soper, S. A. (2022). Microfluidic affinity selection of active SARS-CoV-2 virus particles. Science advances, 8 (39), eabn9665. https://doi.org/10.1126/sciadv.abn9665