Electrochemical DNA Biosensor for detection of Avian Infectious Bronchitis virus (IBV) using Gold Modified Electrode with Glutaraldehyde as cross linker

Abstract

Abstract Infectious bronchitis virus (IBV) belongs to the Gamma coronavirus family of avian species, which is characterized by respiratory infections, nephropathy, and permanent damage to the oviduct, resulting in high mortality in both layer and broiler chickens. IBV has caused significant financial losses and a lack of on-farm diagnostic tools, posing a significant challenge for disease prevention and control. The development of DNA biosensors has increased the importance of rapid and accurate detection of the viral disease in poultry. In this study, an electrochemical DNA biosensor was developed based on a gold electrode fabricated with a nanocomposite of chitosan (CS), multi-walled carbon nanotubes (MWNTs), and highly reactive glutaraldehyde (GLU). The biosensor utilized the target-specific Orf gene of IBV. Under optimal parameters, immobilization and hybridization efficiencies were evaluated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), with methylene blue used as the redox indicator. The proposed DNA biosensor successfully detected the target DNA in a range of 2.0 x 10–12 to 2.0 x 10− 5 molL-1, with a limit of detection (LOD) and limit of quantitation (LOQ) of 2.6 nM and 0.79 nM, respectively. Cross-reactivity studies were performed against non-IBV viruses, consistently validating its sensitivity for detection. The hybridization event was analyzed in real samples by applying the selected probe to the modified gold electrode, using crude cDNA and PCR products. The recovery rate ranged from 95.41–104.55%. These results demonstrate that the proposed DNA biosensor has the potential to accurately detect various strains of IBV in real samples. Additionally, it offers a rapid, sensitive, and alternative technique for diagnosing IBV disease, addressing the current challenges in disease prevention and control.