Comparative Analysis of Hepatitis C Virus (HCV) Screening Tests and Confirmatory PCR: Evaluation of Diagnostic Accuracy, Weak Positive Results, and Clinical Impact

Abstract

Background: Hepatitis C virus (HCV) is a major global public health concern and a leading cause of chronic liver disease. Early and accurate diagnosis is essential to prevent disease progression and transmission. Although rapid diagnostic tests (RDTs) and enzyme-linked immunosorbent assay (ELISA) are widely used for HCV screening, polymerase chain reaction (PCR) remains the gold standard for confirming active infection.

Objective: To compare the diagnostic performance of HCV screening tests (RDT and ELISA) with confirmatory PCR, evaluate their diagnostic accuracy, assess the significance of weakly reactive screening results, and determine the role of PCR in accurate HCV diagnosis and patient management.

Methods: A cross-sectional comparative diagnostic study was conducted from October to December 2025 in diagnostic laboratories of District Okara and District Kasur, Pakistan. A total of 300 participants were enrolled. Serum samples were tested using RDT and ELISA, while PCR was performed as the confirmatory reference test for positive and weakly positive screening samples. Diagnostic performance was assessed using descriptive statistics and standard diagnostic accuracy measures.

Results: Among the 300 participants, PCR confirmed 150 (50.0%) as true positive and 100 (33.3%) as true negative cases. Fifty (16.7%) participants showed weakly reactive or discordant screening results requiring PCR confirmation. The findings demonstrated that RDT and ELISA are effective screening tools; however, weakly reactive results may represent early infection or false-positive reactions and require molecular confirmation. PCR provided the highest diagnostic accuracy for confirming active HCV infection.

Conclusion: RDT and ELISA are suitable for initial HCV screening because of their practicality and sensitivity. However, all positive and weakly reactive screening results should be confirmed by PCR to accurately identify active HCV infection, reduce misdiagnosis, and support appropriate clinical management