Affinity enrichment of extracellular vesicles from plasma reveals mRNA changes associated with acute ischemic stroke.

ABSTRACT: Currently there is no in vitro diagnostic test for acute ischemic stroke (AIS), yet rapid diagnosis is crucial for effective thrombolytic treatment. We previously demonstrated the utility of CD8(+) T-cells' mRNA expression for AIS detection; however extracellular vesicles (EVs) were not evaluated as a source of mRNA for AIS testing. We now report a microfluidic device for the rapid and efficient affinity-enrichment of CD8(+) EVs and subsequent EV's mRNA analysis using droplet digital PCR (ddPCR). The microfluidic device contains a dense array of micropillars modified with anti-CD8? monoclonal antibodies that enriched 158?±?10?nm sized EVs at 4.3 ± 2.1 × 109 particles/100?µL of plasma. Analysis of mRNA from CD8(+) EVs and their parental T-cells revealed correlation in the expression for AIS-specific genes in both cell lines and healthy donors. In a blinded study, 80% test positivity for AIS patients and controls was revealed with a total analysis time of 3.7?h.

SUBMITTER: Wijerathne H 

PROVIDER: S-EPMC7589468 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature