An adaptable platform for in-house hepatitis C serology

Area of Science:

• * Infectious disease diagnostics
• * Molecular biology and immunology
• * Public health and epidemiology

Background:

• * Serological assays are crucial for infectious disease diagnosis and surveillance.
• * Limited availability and high cost of commercial assays hinder large-scale testing, especially for neglected diseases in low and middle-income countries (LMICs).
• * There is a need for affordable and adaptable diagnostic tools.

Purpose of the Study:

• * To develop and validate a cost-effective, adaptable enzyme-linked immunoassay (ELISA) for infectious disease diagnosis.
• * To use hepatitis C virus (HCV) as a proof-of-concept for the developed assay methodology.
• * To demonstrate the potential of this method for diagnosing neglected diseases.

Main Methods:

• * Development of a novel ELISA by combining maltose-binding-protein with a multiepitope HCV protein.
• * Optimization of the assay for high protein concentration and downstream applications.
• * Validation of the assay using human samples from Canada, Denmark, and Gabon, comparing results with a commercial protein.

Main Results:

• * Achieved high sensitivity (98%) and specificity (97%) for the developed ELISA after optimization and accounting for non-specific binding.
• * Successfully produced a high concentration of the engineered protein.
• * Demonstrated the assay's effectiveness using previously tested human samples.

Conclusions:

• * A robust, adaptable, and low-cost multiepitope ELISA for HCV diagnosis has been successfully developed and validated.
• * The methodology is readily transferable for developing diagnostic assays for other infectious pathogens with known immunogenic epitopes.
• * This approach has the potential to significantly improve the diagnosis of neglected diseases in resource-limited settings.