Microfluidic Immunoarray for Point-of-Care Detection of Cytokines in COVID-19 Patients

Area of Science:

• Biomedical Engineering
• Immunology
• Infectious Diseases

Background:

• COVID-19 cytokine storm leads to ARDS, multiorgan failure, and death.
• Elevated pro-inflammatory cytokines correlate with COVID-19 severity.
• Rapid cytokine analysis is crucial for timely treatment and improved patient outcomes.

Purpose of the Study:

• To develop a sensitive, low-cost, semiautomated microfluidic immunoarray for rapid detection of inflammation biomarkers.
• To simultaneously analyze cytokines and CRP in small serum volumes for COVID-19 severity assessment.
• To validate the diagnostic value of the developed system against established methods and disease severity scores.

Main Methods:

• Fabrication of a 3D-printed microfluidic device with immobilized antibodies in microwells.
• Automated reagent delivery using a programmable syringe pump.
• Chemiluminescence signal detection via CCD camera for simultaneous IL-6, IFN-γ, and CRP quantification.
• Validation using 80 COVID-19 patient serum samples and comparison with a commercial immunoassay.

Main Results:

• Achieved ultralow limits of detection: 0.79 fg/mL (IL-6), 4.2 fg/mL (CRP), 2.7 fg/mL (IFN-γ).
• Demonstrated high sensitivity and accuracy, correlating well with a commercial immunoassay.
• ROC analysis indicated good diagnostic value for predicting COVID-19 severity, particularly for IL-6 and CRP.

Conclusions:

• The developed microfluidic immunoarray offers a rapid, cost-effective tool for monitoring COVID-19 severity biomarkers.
• Simultaneous detection of IL-6, IFN-γ, and CRP can aid in predicting disease progression.
• This technology supports critical treatment decisions, potentially improving patient survival rates.