tetano
Editor, Senior Moderator
Sci Adv
. 2022 Sep 30;8(39):eabn9665.
doi: 10.1126/sciadv.abn9665. Epub 2022 Sep 28.
Microfluidic affinity selection of active SARS-CoV-2 virus particles
Sachindra S T Gamage[SUP] 1 2 [/SUP], Thilanga N Pahattuge[SUP] 1 2 [/SUP], Harshani Wijerathne[SUP] 1 2 [/SUP], Katie Childers[SUP] 2 3 [/SUP], Swarnagowri Vaidyanathan[SUP] 2 3 [/SUP], Uditha S Athapattu[SUP] 1 2 [/SUP], Lulu Zhang[SUP] 2 3 [/SUP], Zheng Zhao[SUP] 1 2 [/SUP], Mateusz L Hupert[SUP] 4 [/SUP], Rolf M Muller[SUP] 4 [/SUP], Judy Muller-Cohn[SUP] 4 [/SUP], Janet Dickerson[SUP] 4 [/SUP], Dylan Dufek[SUP] 4 [/SUP], Brian V Geisbrecht[SUP] 5 [/SUP], Harsh Pathak[SUP] 6 [/SUP], Ziyan Pessetto[SUP] 7 [/SUP], Gregory N Gan[SUP] 8 9 [/SUP], Junseo Choi[SUP] 2 10 [/SUP], Sunggook Park[SUP] 2 10 [/SUP], Andrew K Godwin[SUP] 2 6 9 [/SUP], Malgorzata A Witek[SUP] 1 2 [/SUP], Steven A Soper[SUP] 1 2 3 9 11 [/SUP]
Affiliations
Abstract
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.
. 2022 Sep 30;8(39):eabn9665.
doi: 10.1126/sciadv.abn9665. Epub 2022 Sep 28.
Microfluidic affinity selection of active SARS-CoV-2 virus particles
Sachindra S T Gamage[SUP] 1 2 [/SUP], Thilanga N Pahattuge[SUP] 1 2 [/SUP], Harshani Wijerathne[SUP] 1 2 [/SUP], Katie Childers[SUP] 2 3 [/SUP], Swarnagowri Vaidyanathan[SUP] 2 3 [/SUP], Uditha S Athapattu[SUP] 1 2 [/SUP], Lulu Zhang[SUP] 2 3 [/SUP], Zheng Zhao[SUP] 1 2 [/SUP], Mateusz L Hupert[SUP] 4 [/SUP], Rolf M Muller[SUP] 4 [/SUP], Judy Muller-Cohn[SUP] 4 [/SUP], Janet Dickerson[SUP] 4 [/SUP], Dylan Dufek[SUP] 4 [/SUP], Brian V Geisbrecht[SUP] 5 [/SUP], Harsh Pathak[SUP] 6 [/SUP], Ziyan Pessetto[SUP] 7 [/SUP], Gregory N Gan[SUP] 8 9 [/SUP], Junseo Choi[SUP] 2 10 [/SUP], Sunggook Park[SUP] 2 10 [/SUP], Andrew K Godwin[SUP] 2 6 9 [/SUP], Malgorzata A Witek[SUP] 1 2 [/SUP], Steven A Soper[SUP] 1 2 3 9 11 [/SUP]
Affiliations
- PMID: 36170362
- DOI: 10.1126/sciadv.abn9665
Abstract
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.