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Updated: Aug 27, 2025

In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses

In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses

Published on: September 7, 2022

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

Sachindra S T Gamage^1,2, Thilanga N Pahattuge^1,2, Harshani Wijerathne^1,2

¹Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.

Science Advances

|September 28, 2022

Summary

This study introduces a microfluidic assay to isolate active SARS-CoV-2 viral particles from patient samples. The assay demonstrates high sensitivity and specificity for detecting infectious severe acute respiratory syndrome coronavirus 2.

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A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice

A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice

Published on: November 23, 2015

Area of Science:

Biotechnology
Virology
Microfluidics

Background:

Accurate detection of infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial for managing coronavirus disease 2019 (COVID-19).
Current diagnostic methods may not always distinguish between infectious viral particles and non-infectious viral material.
There is a need for sensitive and specific assays to identify active SARS-CoV-2 viral particles (VPs).

Purpose of the Study:

To develop and validate a microfluidic assay for the selection and quantification of active SARS-CoV-2 VPs from clinical samples.
To define active VPs as intact particles with accessible angiotensin-converting enzyme 2 receptor binding domain (RBD) on the spike (S) protein.

Main Methods:

Utilized injection-molded microfluidic chips for high-scale production and screening.
Employed surface-bound aptamers targeting the S protein RBD for affinity selection of SARS-CoV-2 VPs.
Released selected VPs using blue light-emitting diode (LED) for subsequent analysis.
Quantified selected VPs using reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Main Results:

Achieved high recovery (~94%) and release efficiency (89%) of SARS-CoV-2 VPs.
Demonstrated high clinical specificity (100%) in identifying healthy donors.
Showcased high sensitivity (95%) in detecting SARS-CoV-2 VPs in patients with COVID-19, identifying active VPs in 15 out of 20 patients.
The microfluidic chip demonstrated reprogrammability for selecting other viral particles or exosomes by changing the affinity agent.

Conclusions:

The developed microfluidic assay effectively selects and enumerates active SARS-CoV-2 viral particles from clinical samples.
The assay offers high sensitivity and specificity, aiding in the diagnosis of COVID-19 by detecting infectious viral particles.
The adaptable design of the microfluidic chip holds potential for broader applications in pathogen detection and exosome analysis.