Development of in vitro diagnostic (IVD) kits against hepatitis A and E infections

Abstract

Viral hepatitis, characterized by hepatic inflammation, is predominantly caused by hepatoviruses such as Hepatitis A virus (HAV) and Hepatitis E virus (HEV). This study aims to develop a rapid, in vitro diagnostic tool for detecting HAV and HEV infections using an antigen-based Lateral Flow Immunoassay (LFIA). The proposed diagnostic kit utilizes recombinant capsid proteins of HAV and HEV, which serve as specific antigens to capture anti-HAV/HEV IgM and IgG antibodies from patient serum samples. These antigens were produced by cloning the respective capsid protein genes into ‘Escherichia coli’ expression systems, followed by purification and validation through enzyme-linked immunosorbent assay (ELISA) to confirm antigenicity. Optimization of the ELISA protocol enabled clear serological differentiation between infected and uninfected human sera. Crossreactivity studies confirmed high specificity of the antigens, with negligible response against non-target hepatitis viruses. Statistical analysis further substantiated significant differences in optical density (O.D.) values between target-specific and non-specific interactions. Development of the LFIA device is ongoing, with current efforts focused on optimizing assay sensitivity, visual clarity, and consistency in differentiating positive and negative results. Keywords: Hepatitis A, Hepatitis E, ELISA, LFIA, Diagnostics.