Muhammad Huzaifa, Syed Waqas Hassan
The Hepatitis C virus (HCV) infection is a serious problem for health all around the world. It is especially in low resource settings where people do not have sufficient resources to access medical care. This work developed a test for HCV infection, which analytically assessed an immunochromatographic (ICT) based on gold nanoparticles ((AuNPs) for detection of anti HCV antibodies in human serum samples. The optimization of nanoparticles synthesis by redox method, antibodies and antigen conjugation, and multi-component buffer formulations improved the stability, sensitivity, specificity and signal intensity of the assay system. The effective surface fictionalization, non specific bindings, and critical factors such as pH, protein concentration, blocking conditions, and stabilization buffer were optimized to achieve more sensitive reliable testing method. The developed assay demonstrated high diagnostic performance, with sensitivity up to 97.5%, specificity 98.2% and accuracy 97.6% using clinical samples. The positive predictive value (PPV) and negative predictive value (NPV) were calculated as 97.98% and 97.03% respectively. The assay showed a limit of detection of ELISA S/Co value of approximately 1.14, with excellent linearity (R² = 0.991), confirming its capability for low-level antibody detection. The developed ICT provided rapid results within 10-15 minutes and demonstrated enhanced signal intensity compared to conventional lateral flow systems. Overall, this study presents a robust, scalable, and cost-effective diagnostic platform for HCV screening.