IFN-γ and IL-10 Immunosensor with Vertically Aligned Carbon Nanotube Interdigitated Electrodes toward Pen-Side Cattle Paratuberculosis Monitoring

Abstract

Highly sensitive vertically aligned carbon nanotube arrays (VANTAs) interdigitated electrode (IDE) arrays are developed for electrochemical biosensing of two cytokines (i.e., interleukin-10 (IL-10) and interferon-gamma (IFN-γ)) that are useful for early detection Johne's disease (Bovine Paratuberculosis) in cattle. The high aspect ratio VANTA-IDEs (50–60 µm in height) are grown through a chemical vapor deposition process from an iron (Fe) catalyst that is lithographically patterned on a silicon wafer with equal finger width and inter-finger spacing of 25 µm. After functionalization with distinct antibodies the VANTA-IDEs are capable of selective detection of both IL-10 and IFN-γ within an actual biological matrix (i.e., diluted bovine implant supernatant) over concentration ranges of 0.1 to 30 pg mL⁻¹ (limit of detection – LOD: 0.0911 pg mL⁻¹) and 50–500 pg mL⁻¹ (LOD: 24.17 pg mL⁻¹), respectively with a response time of <35 min. Results demonstrate important initial steps for rapid, pen-side identification of cattle with stage-I Mycobacterium avium subspecies paratuberculosis infection before physical symptoms of Johne's disease are present. Such a rapid pen-side diagnostic test can be used on cattle at an auction or before they are introduced to a herd to ensure the larger population does not become infected with Johne's disease.