Development of rapid colorimetric assay for the detection of Influenza A and B viruses

Abstract

The flu viruses are respiratory pathogens which, according to the World Health Organization (WHO), infect 5–10% of the world population resulting in 3–5 million cases of severe illness and 290,000 to 650,000 annual deaths. Early diagnosis and therapeutic intervention can ameliorate symptoms of infection and reduce mortality. The conventional diagnosis of viral infections, including flu viruses, has evolved over the years with diverse approaches, however, there are inherent short comings associated with these testing. There is an urgent need for rapid and low-cost diagnostic assays, due to the enormous annual burden of influenza diseases and its associated mortality . In this study, novel, low cost and easy to use colorimetric flu virus biosensor assay was developed. The sandwich assay format was utilized using antibodies immobilized onto cotton swabs, for the rapid detection of flu A and B viruses. These swabs serve as sample collection, analytes pre-concentration as well as sensing tool. The proof of concept was established for this assay in buffer and mucus samples. The limit of detection (LOD) of the colorimetric assay was 0.04 ng mL −1 for Flu A and Flu B respectively and with linear dynamic range between 0.04 ng ml −1 to 40 ng ml for both viruses in mucous samples. The assay can be performed at the patient's bed side by minimally skilled hospital personnel without the need for instrumentation. Cross-reactivity assays testing was done using Flu viruses specific activated swabs reacted with other common respiratory viral pathogens' antigen, in order to assess the specificity of the swabs.

Recent advances in the biosensors application for the detection of bacteria and viruses in wastewater

“…In the past couple of years, viral isolation, serology, nucleic amplification, and lab-on-chip assays have been used and modified for Flu viruses' diagnostics. However, due to the common drawbacks of these diagnostic measures and the overwhelming concerns associated with mortality rates, there continues to be a need for rapid diagnostic tools for Flu viruses [52] , [63] . Raji et al reported the development of biosensors for the detection of Flu A and Flu B viruses [52] .…”

“…However, due to the common drawbacks of these diagnostic measures and the overwhelming concerns associated with mortality rates, there continues to be a need for rapid diagnostic tools for Flu viruses [52] , [63] . Raji et al reported the development of biosensors for the detection of Flu A and Flu B viruses [52] . This study used antibodies for Flu A and Flu B viruses and immobilized them on the surface of colorimetric biosensors.…”

Nanotechnology on agent identification, diseases diagnosis and therapeutic approaches

“…These nanoparticles are applied to increase the sensitivity and effectiveness in detecting microorganisms, reducing time, costs, and the need for advanced laboratory structures to perform the diagnosis (Jain et al, 2020;Landry et al, 2017;Li et al, 2018;Safarpour et al, 2021). Immunoagglutination using gold (AuNP) and DNA probes immobilization on surfaces coated with metallic nanoparticles (AuNPs and MNPs) where the DNA probe on the surface of AuNPs was a signal reporter by fluorescence are some examples of this application (Raji et al, 2021).…”

“…The immunoassay test from the association of metallic nanoparticles was reported with monoclonal antibodies that bind conjugately to epitopes (molecules, DNA, cells) of the microorganism on a surface with an affinity to the antibody (Basu et al, 2004;Basso et al, 2018;Mikaelyan et al, 2017). This surface contains antibodies specific for the antibody associated with the nanoparticle, which retains them and allows visualization by colorimetric change (Raji et al, 2021). This association has shown to be a promising platform for outbreak elucidation and diagnosis of various agents (Bhattarakosol, et al, 2018;Chen et al, 2018;Chen et al, 2019;Elahi et al, 2019;Vedova-Costa et al, 2021), as well as monitoring the evolution of neurodegenerative diseases such as Alzheimer's and diabetes (Popli et al, 2018).…”

Development of a Low-Cost Cotton-Tipped Electrochemical Immunosensor for the Detection of SARS-CoV-2

“…Cotton swabs have been recently used in the fabrication of immunoassays for the detection of different pathogens. − In these assays, the colorimetric detection was achieved based on visual discrimination of the color change. These assays are simple, fast, and easy to perform.…”