Detection of occult hepatitis C virus infection among Egyptian patients with hepatocellular carcinoma who achieved sustained virologic response to direct-acting antiviral therapy

Kassem, Fatma E.M.A.; El-Kholy, Emtethal E.; Mohamed, Amira H.; Abdul-Mohaymen, Abeer M.; Gomaa, Asmaa I.; Hassan, Ayat S.M.

doi:10.21608/epj.2025.407585

Detection of occult hepatitis C virus infection among Egyptian patients with hepatocellular carcinoma who achieved sustained virologic response to direct-acting antiviral therapy

Document Type : Original Article

Authors

¹ Department of Electron Microscopy, Theodore Bilharz Research Institute, Giza, Egypt

² Department of Clinical Pathology, Faculty of Medicine for Girls, Al-Azhar University, Cairo, Egypt

³ Department of Tropical Medicine, National Liver Institute, Menoufia University, Menoufia, Egypt

10.21608/epj.2025.407585

Abstract

Background
Occult hepatitis C virus infection (OCI) is defined as the presence of hepatitis C virus (HCV) RNA in nonserum reservoirs without any detectable HCV-RNA in serum by standard assays. The association between OCI and hepatocellular carcinoma (HCC) is controversial. Direct-acting antiviral (DAA) therapy effectively controls HCV infection and proves sustained virological response (SVR). However, HCC after DAA therapy is debatable.
Objective
In this study, we tried to detect OCI in HCC Egyptian patients who achieved SVR post-DAA therapy for possible correlation and prediction for HCC.
Patients and methods
A cross–sectional study included seventy HCC Egyptian patients who achieved SVR post sofosbuvir/daclatasvir (SOF/DACLA) +/–ribavirin therapy. HCV-RNA was detected in peripheral blood mononuclear cells (PBMCs) by RT-PCR. Immune transmission electron microscopy (TEM) was done to study OCI in PBMCs using specific anti-HCV antibodies.
Results and conclusion
TEM examination revealed positive OCI in PBMCs of all patients (70/70) with 3 stages of grading reflecting viral particle load in PBMCs, with 68.6% of the patients having a marked OCI grade, while PCR only detected 5.7% (4/70) of OCI. Nearly 67.1% of the cases received dual anti-HCV therapy (sofosbuvir/daclatasvir). Ribavirin-inclusive DAA treatment shows a higher grade of OCI (P=0.02). The average duration from SVR to HCC development was 29.8±13.4 months. In conclusion, DAA drugs effectively eliminate HCV in serum, but OCI is still considered a risk for developing HCC, recommending a re-definition for SVR through detecting HCV-RNA in serum, PBMCs, and TEM studies. Our study is the first to provide electron microscopy as a sensitive tool for OCI detection with established superiority over the PCR technique.

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