Multiplexed biosensor for point-of-care COVID-19 monitoring: CRISPR-powered unamplified RNA diagnostics and protein-based therapeutic drug management

Abstract

In late 2019 SARS-CoV-2 rapidly spread to become a global pandemic, therefore, measures to attenuate chains of infection, such as high-throughput screenings and isolation of carriers were taken. Prerequisite for a reasonable and democratic implementation of such measures, however, is the availability of sufficient testing opportunities (beyond reverse transcription PCR, the current gold standard). We, therefore, propose an electrochemical, microfluidic multiplexed biosensor in combination with CRISPR/Cas-powered assays for point-of-care nucleic acid testing. In this study, we simultaneously screen for and identify SARS-CoV-2 infections (Omicron-variant) in clinical specimens (Sample-to-result time: ∼30 min), employing LbuCas13a, whilst bypassing reverse transcription as well as target amplification of the viral RNA, both of which are necessary for detection via PCR and multiple other methods. In addition, we demonstrate the feasibility of combining assays based on different classes of biomolecules, in this case protein-based antibiotic detection, on the same device. The programmability of the effector and multiplexing capacity (up to six analytes) of our platform, in combination with a miniaturized measurement setup, including a credit card sized near field communication (NFC) potentiostat and a microperistaltic pump, provide a promising on-site tool for identifying individuals infected with variants of concern and monitoring their disease progression alongside other potential biomarkers or medication clearance.

Designing electrochemical microfluidic multiplexed biosensors for on-site applications

“…(iii) Using the BiosensorX platform in clinical applications, the measurement setup needs to be further miniaturized, automated, and/or easier to use. Herein, a smartphone-compatible small potentiostat for signal readout along with a micropump for the stop-flow protocol could be used [38]. The BiosensorX presented in this work is capable of detecting several analytes simultaneously and can be further extended to detect several biomarkers and anti-infective agents in different sample types on a single chip, which is a huge step towards fast and easy therapeutic drug management at the point care.…”

Smartphone-Based Multiplexed Biosensing Tools for Health Monitoring

“…Biomarkers can be used to objectively assess pathogenic processes since both qualitative and quantitative detection is crucial for disease identification [ 107 ]. Viruses, such as the highly pathogenic Asian avian (H5N1) influenza A, Zika, Ebola, hepatitis B and hepatitis C, coronaviruses (CoVs), SARS-CoV-1 and SARS-CoV-2, and Middle East respiratory syndrome (MERS-CoV), cause a variety of infectious diseases that affect millions of people [ 49 , 108 , 109 , 110 ]. However, the identification of a single biomarker is insufficient to identify a disease.…”

NFC Smartphone-based electrochemical microfluidic device integrated with nanobody recognition for C-reactive protein

“…Its primary advantage is the ability to provide real-time analysis, enabling healthcare providers to make immediate treatment decisions and improve patient outcomes 1,2 . With the growing demand for personalized and timely healthcare, as evidenced during the COVID-19 pandemic, the role of POCT continues to expand, contributing to more efficient and effective healthcare [3][4][5][6] .…”