Microfluidic affinity selection of active SARS-CoV-2 virus particles.

Autor: Gamage SST; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Pahattuge TN; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Wijerathne H; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Childers K; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Bioengineering Program, The University of Kansas, Lawrence, KS 66045, USA., Vaidyanathan S; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Bioengineering Program, The University of Kansas, Lawrence, KS 66045, USA., Athapattu US; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Zhang L; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Bioengineering Program, The University of Kansas, Lawrence, KS 66045, USA., Zhao Z; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Hupert ML; BioFluidica Inc., San Diego, CA 92121, USA., Muller RM; BioFluidica Inc., San Diego, CA 92121, USA., Muller-Cohn J; BioFluidica Inc., San Diego, CA 92121, USA., Dickerson J; BioFluidica Inc., San Diego, CA 92121, USA., Dufek D; BioFluidica Inc., San Diego, CA 92121, USA., Geisbrecht BV; Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS 66506, USA., Pathak H; Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS 66160, USA., Pessetto Z; Sinochips Diagnostics, Olathe, KS 66061, USA., Gan GN; Department of Radiation Oncology, University of Kansas Medical Center, Kansas City, KS 66160, USA.; University of Kansas Cancer Center, University of Kansas Medical Center, Kansas City, KS 66160, USA., Choi J; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Department of Industrial and Mechanical Engineering, Louisiana State University, Baton Rouge, LA 70803, USA., Park S; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Department of Industrial and Mechanical Engineering, Louisiana State University, Baton Rouge, LA 70803, USA., Godwin AK; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS 66160, USA.; University of Kansas Cancer Center, University of Kansas Medical Center, Kansas City, KS 66160, USA., Witek MA; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA., Soper SA; Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.; Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.; Bioengineering Program, The University of Kansas, Lawrence, KS 66045, USA.; University of Kansas Cancer Center, University of Kansas Medical Center, Kansas City, KS 66160, USA.; Department of Mechanical Engineering, The University of Kansas, Lawrence, KS 66045, USA.
Jazyk: angličtina
Zdroj: Science advances [Sci Adv] 2022 Sep 30; Vol. 8 (39), pp. eabn9665. Date of Electronic Publication: 2022 Sep 28.
DOI: 10.1126/sciadv.abn9665
Abstrakt: 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.
Databáze: MEDLINE