The surface plasmon resonance biosensor capability for detection avian influenza H5N1 in poultry using immunoglobulin yolk as bioreceptor

• The producing immunoglobulin yolk anti-avian influenza H5N1using hyperimmunization with inactivated vaccine methods in chickens are characterized with SDS-PAGE. • The SPR system is using a nanoSPR8 device with a gold nanoparticle chip coated by IgY anti-avian influenza H5N1 for antigenic detection of H5N1 virus. • The developed SPR biosensor using IgY anti-H5N1 at a concentration of 50 µg/mL demonstrated a limit of detection of 2.24 HAU, with a response sensitivity of 3.90 ΔRU/Log HAU. • The SPR responses' specificity and selectivity to detect AI H5N1 show a significantly different response based on the ΔRU values obtained. • The positive H5N1 from field samples was conducted in the SPR system and showed a significant response. Avian influenza is a viral infectious disease that causes high mortality and morbidity in poultry. The highly pathogenic avian influenza (HPAI) threat causes considerable losses to broiler and laying hens. Early detection of avian influenza cases on farms is now difficult, especially in laying hens with symptoms of decreased egg production, which could be caused by various other diseases. The detection of avian influenza, according to WOAH recommendations, utilizes real-time RT-PCR. The RT-PCR methods are costly, require sample extraction and laboratory expertise. Not only are sensitive and specific alternative diagnostic methods required to address the future threat of avian influenza in poultry with large populations, but also producing biomaterial as a bioreceptor on a large scale is required. Immunoglobulin yolk (IgY) is an antibody produced by the hen and found in egg yolk. Producing IgY requires hyperimmune chickens obtained through the vaccination process. The ability of IgY to bind to antigens has the potential to serve as a bioreceptor for disease detection methods in biosensor devices. A surface plasmon resonance (SPR) biosensor can detect interactions between two molecules, such as antibody-antigens, indicating a change in the resonance angle as a response unit (r.u.). The purpose of this study is to investigate the capability of the SPR biosensor for the diagnosis of AI H5N1 through its limit of detection and sensitivity using IgY anti-avian influenza H5N1, as well as the SPR specificity response in detecting avian influenza in field sample simulations. SPR development using the IgY anti-avian influenza H5N1 bioreceptor with a concentration of 50 µg/ml has a detection limit value of virus titer AI H5N1 10 4,4 ELD50/ml and a response sensitivity of 3.90 ΔRU/Log HAU with an R 2 value of the linear plot from antigen concentration 1 - 16 HAU is 0.99052. IgY anti-avian influenza H5N1 with a concentration of 50 µg/ml in nanoSPR8 device has a reasonable specificity of 89% and selectivity in capturing H5N1 analyte targets. The SPR results are also promising for AI H5N1 rapid detection without extraction due to its positive response in field samples that confirmed AI H5N1 by RT-PCR.