High prevalence of seronegative occult hepatitis C in high-risk individuals
-
Azadeh Haghi Navand
,
Azarakhsh Azaran
,
Mina Bavi
,
Farid Yousefi
,
Maryam Dastorpour
,
Manoochehr Makvandi
Abstract
Background and Objectives: Routine screening for occult hepatitis C virus (OHCV) is not a standard procedure in medical laboratories, which has resulted in an increased incidence of OHCV among high-risk groups and the general population. The objective of this study was to investigate the molecular epidemiology of (OHCV) in Iranian injecting drug users (IDUs).
Materials and Methods: To determine chronic hepatitis C virus(HCV) and OHCV, plasma and peripheral blood mononuclear cell (PBMC) were collected from 103 (96 (93.2%) males, 7 (6.79%) females) IDUs. Their plasma was tested for Anti-HCV (ELISA). Following RNA extraction from plasma and PBMCs, RT-nested PCR was employed to amplify the 5′ untranslated region (5′UTR) and core regions of the HCV genome in plasma and PBMCs from IDUs. Sequencing of the 5′UTR and core regions, along with phylogenetic tree construction, was used to determine HCV genotypes.
Results: Among the 103 individuals, 12/96 males (12.5%) were positive for both anti-HCV and HCV RNA in plasma, indicating chronic HCV infection. In addition, 18/96 males (18.75%) and 1/7 females (14.28%) were positive for anti-HCV but negative for HCV RNA, indicating evidence of past HCV infection (p = 0.1). Furthermore, 5 individuals, including 4/94 males (4.1%) and 1/7 females (14.28%), were found to be seropositive for HCV (p = 0.77). Meanwhile, 23/103 individuals (22.33%), including 20/96 males (20.8%) and 3/7 females (42.85%), were seronegative for HCV (p = 0.37). HCV genotype 1a was the dominant genotype among IDUs.
Conclusion: In conclusion, a high prevalence of HCV infection was observed among IDUs, underscoring the pressing necessity for the implementation of an efficacious strategy to eradicate HCV transmission in this high-risk population.
1. World Health Organization. Hepatitis C. Available online: https://www.who.int/news-room/fact-sheets/detail/hepatitis-c (accessed on 25 July 2025).
2. Hasanvand B, Mirzaei H, Hadian B, Sasaei P, Garavand S, Adhikary H, et al. Prevalence of occult hepatitis C virus infection (OCI) among hemodialysis patients; a cross-sectional study from Lorestan Province, Western Iran. Virusdisease 2024; 35: 478-483.
3. Lanièce Delaunay C, Cox J, Klein M, Lambert G, Grace D, Lachowsky NJ, et al. Trends in hepatitis C virus seroprevalence and associated risk factors among men who have sex with men in Montréal: results from three cross-sectional studies (2005, 2009, 2018). Sex Transm Infect 2021; 97: 290-296.
4. Le DHH, Kanokudom S, Nguyen HM, Yorsaeng R, Honsawek S, Vongpunsawad S, et al. Hepatitis C Virus-Core Antigen: Implications in Diagnostic, Treatment Monitoring and Clinical Outcomes. Viruses 2024; 16: 1863.
5. El Hennawy HM, Abdelaziz AA, Mansour Y, Zaitoun MF, Tawhari I, Safar O, et al. Silent Threat: Fatal Consequences of Occult Hepatitis C Viral Infection in a Kidney Transplant Recipient: A Case Report and Literature Review. Transplant Proc 2025; 57: 445-449.
6. Alshwayyat S, Hanifa H, Almasri N, Taimeh Z, Alshwaiyat Y, Khdair T, et al. Hepatitis C virus among hemodialysis patients: burden, risks, and prevention in resource-limited regions. Int Urol Nephrol 2025;57:3725-3731.
7. Uribe-Noguez LA, Prieto-Torres ME, Uribe-Noguez LO, Mata-Marín JA, Arroyo-Anduiza CI, Paquentín-Jimenez R, et al. Prevalence and Risk Factors of Occult HCV Infection in the Adult Population of Mexico City. Viruses 2025; 17: 236.
8. Tantuoyir MM, Camara M, Sohrabi M, SeyedAlinaghi S, Ahmadinejad Z. Hepatitis C virus infection in Iran: A review of current prevalence and preventive strategies from a developing country. J Clin Virol 2024; 4: 100186.
9. Curado A, Nogueira PJ, Virgolino A, Santa Maria J, Mendão L, Furtado C, et al. Hepatitis C antibody prevalence and behavioral correlates in people who inject drugs attending harm reduction services in Lisbon, Portugal. Front Public Health 2022; 10: 952909.
10. Fleurence RL, Alter HJ, Collins FS, Ward JW. Global elimination of hepatitis C virus. Annu Rev Med 2025; 76: 29-41.
11. Makvandi M, Azaran A, Jalilian S, Nahad MP, Azizolahi B, Neisi N. Frequency and genotype distribution of hepatitis C virus infection in patients with diabetes type 2 in Ahvaz, Iran. Iran J Microbiol 2021; 13: 703 -711.
12. Sayad B, Bozorgomid A, Sayad N, Azhdari M, Bahadori M, Rezaeian S, Gholizadeh M. The prevalence of hepatitis C virus genotypes and factors associated with cirrhosis, fatty liver, and viral load: A registry‐based cross‐sectional cohort study in Western Iran during 1999–2023. Health Sci Rep 2024;7(9): e70079.
13. Tarahomi M, Eslami M, Rad RH, Pooya P, Habibian F, Neyestanaki MH, et al. Hepatitis B and hepatitis C infections in the City of Semnan between 2021 and 2022: A seroepidemiological research. Iran Red Crescent Med J 2024; 45: 1-7.
14. Najimi-Varzaneh A, Gholami-Fesharaki M. Prevalence and distribution of Hepatitis C virus in Iranian drug user: systematic review and meta-analysis study. J Biostat Epidemiol 2020; 6: 72-80.
15. Salari N, Kazeminia M, Hemati N, Ammari-Allahyari M, Mohammadi M, Shohaimi S. Global prevalence of hepatitis C in general population: A systematic review and meta-analysis. Travel Med Infect Dis 2022; 46: 102255.
16. Shiffman ML. The next wave of hepatitis C virus: The epidemic of intravenous drug use. Liver Int 2018; 38(Suppl 1): 34-39.
17. Khezri M, Mirzazadeh A, Shokoohi M, Sharafi H, Ghalekhani N, Tavakoli F, et al. Hepatitis C virus prevalence, determinants, and cascade of care among people who inject drugs in Iran. Drug Alcohol Depend 2023; 243: 109751.
18. Shayan SJ, Nazari R, Kiwanuka F. Prevalence of HIV and HCV among injecting drug users in three selected WHO-EMRO countries: a meta-analysis. Harm Reduct J 2021; 18: 59.
19. Aghaei AM, Gholami J, Sangchooli A, Rostam-Abadi Y, Olamazadeh S, Ardeshir M, et al. Prevalence of injecting drug use and HIV, hepatitis B, and hepatitis C in people who inject drugs in the Eastern Mediterranean region: a systematic review and meta-analysis. Lancet Glob Health 2023; 11(8): e1225-e1237.
20. Karimi SE, Bayani A, Higgs P, Bayat AH, Hemmat M, Ahounbar E, et al. Prevalence and high risk behaviors associated with HCV testing among people who inject drugs: a systematic review and Meta-analysis. Subst Abuse Treat Prev Policy 2020;15: 64.
21. Silva E, Marques S, Leal B, Canhao B, Madaleno J, Simao A, et al. Occult hepatitis C infection identified in injection drug users with direct antiviral agents therapy and spontaneous resolution of hepatitis C virus infection. Virus Res 2023; 329: 199104.
22. Elkashef KA, Fattah NR, Mesbah NM, El-Shaarawy FF, Amer M, Fakhr AE, Gharib AF, Abo-Elmatty DM, Abdel-Hamed AR. The hidden epidemic of occult hepatitis B and C among injection drug users (IDUs): A call for action. Int J Health Sci (Qassim) 2025; 19: 22-30.
23. Naguib H, Abouelnaga SF, Elsayed MM. Occult hepatitis C virus infection in hemodialysis patients who achieved a sustained virological response to directly acting antiviral drugs: is it a concern?. Int Urol Nephrol 2024; 56: 217-222.
24. Chang M, Chen W, Pao L, Chiu C, Chien R. HCV Ab titer and ALT level indicate occult hepatitis C virus infection in treatment-naive HCV Ab-positive and HCV Ab-negative patients: a 3-year prospective cohort study. Microbiol Spectr 2025; 13(8): e0292224.
25. Lotfi AA, Mohamed AE, Shalaby NA, Eissa DS, El-Dabaa E, Sallam AM, et al. Occult hepatitis C virus infection in patients with malignant lymphoproliferative disorders. Int J Immunopathol Pharmacol 2020; 34: 2058738420961202.
26. Azzam A, Khaled H, Hussein SM, Gendy KR, Hassan FE. Prevalence and clinical implications of occult hepatitis C infection: a meta-analysis of studies conducted in Egypt. Egyp J Intern Med 2025; 37: 45.
27. Yaghobi R, Kazemi MJ, Geramizadeh B, Malek Hosseini SA, Moayedi J. Significance of occult hepatitis C virus infection in liver transplant patients with cryptogenic cirrhosis. Exp Clin Transplant 2020; 18: 206-209.
28. Khattab MA, Zakaria Y, Sadek E, Abd El Fatah AS, Fouad M, Khattab M, et al. Detection of hepatitis C virus (HCV) RNA in the peripheral blood mononuclear cells of HCV-infected patients following sustained virologic response. Clin Exp Med 2023; 23: 131-140.
29. Saad ZM, Ghany WAE, Khalifa R, Higazi A, Al-Shazly M, Said M, et al. Occult HCV infection in liver transplanted patients: frequency and consequences. Clin Exp Hepatol 2022; 8: 125-131.
30. Namayandeh M, Jamalidoust M, Heydari Marandi N, Aliabadi N, Ziyaeyan A, Pouladfar G, et al. Hepatitis C virus genotypes in patients with chronic hepatitis C infection in southern Iran from 2016 to 2019. Microbiol Immunol 2020; 64: 762-767.
31. Keikha M, Eslami M, Yousefi B, Ali-Hassanzadeh M, Kamali A, Yousefi M, et al. HCV genotypes and their determinative role in hepatitis C treatment. Virusdisease 2020; 31: 235-240.
32. Tsai MC, Hung CH, Lu SN, Wang JH, Chen CH, Kee KM, et al. The incidence of resistance-associated variants to NS5A in HCV subtypes 1a and 1b in Taiwan. Biomed J 2021; 44(6 Suppl 1): S126-S131.
33. Ryan P, Valencia J, Cuevas G, Amigot-Sanchez R, Martínez I, Lazarus JV, et al. Decrease in active hepatitis C infection among people who use drugs in Madrid, Spain, 2017 to 2023: a retrospective study. Euro Surveill 2024; 29: 2300712.
34. Scarpetta M, Kanner R, Menezes NP, McDonell CC, Bruneau J, Page K, et al. Getting to full disclosure: HCV testing and status disclosure behaviors among PWID and their injecting partners. BMC Public Health 2025; 25: 1707.
35. Goel A, Rewari BB, Sharma M, Konath NM, Aggarwal R. Seroprevalence and burden of hepatitis C virus infection in WHO South-East Asia Region: A systematic review. J Gastroenterol Hepatol 2022;37:964-972.
36. Torre P, Festa M, Sarcina T, Masarone M, Persico M. Elimination of HCV Infection: Recent Epidemiological Findings, Barriers,and Strategies for the Coming Years. Viruses 2024; 16: 1792.
2. Hasanvand B, Mirzaei H, Hadian B, Sasaei P, Garavand S, Adhikary H, et al. Prevalence of occult hepatitis C virus infection (OCI) among hemodialysis patients; a cross-sectional study from Lorestan Province, Western Iran. Virusdisease 2024; 35: 478-483.
3. Lanièce Delaunay C, Cox J, Klein M, Lambert G, Grace D, Lachowsky NJ, et al. Trends in hepatitis C virus seroprevalence and associated risk factors among men who have sex with men in Montréal: results from three cross-sectional studies (2005, 2009, 2018). Sex Transm Infect 2021; 97: 290-296.
4. Le DHH, Kanokudom S, Nguyen HM, Yorsaeng R, Honsawek S, Vongpunsawad S, et al. Hepatitis C Virus-Core Antigen: Implications in Diagnostic, Treatment Monitoring and Clinical Outcomes. Viruses 2024; 16: 1863.
5. El Hennawy HM, Abdelaziz AA, Mansour Y, Zaitoun MF, Tawhari I, Safar O, et al. Silent Threat: Fatal Consequences of Occult Hepatitis C Viral Infection in a Kidney Transplant Recipient: A Case Report and Literature Review. Transplant Proc 2025; 57: 445-449.
6. Alshwayyat S, Hanifa H, Almasri N, Taimeh Z, Alshwaiyat Y, Khdair T, et al. Hepatitis C virus among hemodialysis patients: burden, risks, and prevention in resource-limited regions. Int Urol Nephrol 2025;57:3725-3731.
7. Uribe-Noguez LA, Prieto-Torres ME, Uribe-Noguez LO, Mata-Marín JA, Arroyo-Anduiza CI, Paquentín-Jimenez R, et al. Prevalence and Risk Factors of Occult HCV Infection in the Adult Population of Mexico City. Viruses 2025; 17: 236.
8. Tantuoyir MM, Camara M, Sohrabi M, SeyedAlinaghi S, Ahmadinejad Z. Hepatitis C virus infection in Iran: A review of current prevalence and preventive strategies from a developing country. J Clin Virol 2024; 4: 100186.
9. Curado A, Nogueira PJ, Virgolino A, Santa Maria J, Mendão L, Furtado C, et al. Hepatitis C antibody prevalence and behavioral correlates in people who inject drugs attending harm reduction services in Lisbon, Portugal. Front Public Health 2022; 10: 952909.
10. Fleurence RL, Alter HJ, Collins FS, Ward JW. Global elimination of hepatitis C virus. Annu Rev Med 2025; 76: 29-41.
11. Makvandi M, Azaran A, Jalilian S, Nahad MP, Azizolahi B, Neisi N. Frequency and genotype distribution of hepatitis C virus infection in patients with diabetes type 2 in Ahvaz, Iran. Iran J Microbiol 2021; 13: 703 -711.
12. Sayad B, Bozorgomid A, Sayad N, Azhdari M, Bahadori M, Rezaeian S, Gholizadeh M. The prevalence of hepatitis C virus genotypes and factors associated with cirrhosis, fatty liver, and viral load: A registry‐based cross‐sectional cohort study in Western Iran during 1999–2023. Health Sci Rep 2024;7(9): e70079.
13. Tarahomi M, Eslami M, Rad RH, Pooya P, Habibian F, Neyestanaki MH, et al. Hepatitis B and hepatitis C infections in the City of Semnan between 2021 and 2022: A seroepidemiological research. Iran Red Crescent Med J 2024; 45: 1-7.
14. Najimi-Varzaneh A, Gholami-Fesharaki M. Prevalence and distribution of Hepatitis C virus in Iranian drug user: systematic review and meta-analysis study. J Biostat Epidemiol 2020; 6: 72-80.
15. Salari N, Kazeminia M, Hemati N, Ammari-Allahyari M, Mohammadi M, Shohaimi S. Global prevalence of hepatitis C in general population: A systematic review and meta-analysis. Travel Med Infect Dis 2022; 46: 102255.
16. Shiffman ML. The next wave of hepatitis C virus: The epidemic of intravenous drug use. Liver Int 2018; 38(Suppl 1): 34-39.
17. Khezri M, Mirzazadeh A, Shokoohi M, Sharafi H, Ghalekhani N, Tavakoli F, et al. Hepatitis C virus prevalence, determinants, and cascade of care among people who inject drugs in Iran. Drug Alcohol Depend 2023; 243: 109751.
18. Shayan SJ, Nazari R, Kiwanuka F. Prevalence of HIV and HCV among injecting drug users in three selected WHO-EMRO countries: a meta-analysis. Harm Reduct J 2021; 18: 59.
19. Aghaei AM, Gholami J, Sangchooli A, Rostam-Abadi Y, Olamazadeh S, Ardeshir M, et al. Prevalence of injecting drug use and HIV, hepatitis B, and hepatitis C in people who inject drugs in the Eastern Mediterranean region: a systematic review and meta-analysis. Lancet Glob Health 2023; 11(8): e1225-e1237.
20. Karimi SE, Bayani A, Higgs P, Bayat AH, Hemmat M, Ahounbar E, et al. Prevalence and high risk behaviors associated with HCV testing among people who inject drugs: a systematic review and Meta-analysis. Subst Abuse Treat Prev Policy 2020;15: 64.
21. Silva E, Marques S, Leal B, Canhao B, Madaleno J, Simao A, et al. Occult hepatitis C infection identified in injection drug users with direct antiviral agents therapy and spontaneous resolution of hepatitis C virus infection. Virus Res 2023; 329: 199104.
22. Elkashef KA, Fattah NR, Mesbah NM, El-Shaarawy FF, Amer M, Fakhr AE, Gharib AF, Abo-Elmatty DM, Abdel-Hamed AR. The hidden epidemic of occult hepatitis B and C among injection drug users (IDUs): A call for action. Int J Health Sci (Qassim) 2025; 19: 22-30.
23. Naguib H, Abouelnaga SF, Elsayed MM. Occult hepatitis C virus infection in hemodialysis patients who achieved a sustained virological response to directly acting antiviral drugs: is it a concern?. Int Urol Nephrol 2024; 56: 217-222.
24. Chang M, Chen W, Pao L, Chiu C, Chien R. HCV Ab titer and ALT level indicate occult hepatitis C virus infection in treatment-naive HCV Ab-positive and HCV Ab-negative patients: a 3-year prospective cohort study. Microbiol Spectr 2025; 13(8): e0292224.
25. Lotfi AA, Mohamed AE, Shalaby NA, Eissa DS, El-Dabaa E, Sallam AM, et al. Occult hepatitis C virus infection in patients with malignant lymphoproliferative disorders. Int J Immunopathol Pharmacol 2020; 34: 2058738420961202.
26. Azzam A, Khaled H, Hussein SM, Gendy KR, Hassan FE. Prevalence and clinical implications of occult hepatitis C infection: a meta-analysis of studies conducted in Egypt. Egyp J Intern Med 2025; 37: 45.
27. Yaghobi R, Kazemi MJ, Geramizadeh B, Malek Hosseini SA, Moayedi J. Significance of occult hepatitis C virus infection in liver transplant patients with cryptogenic cirrhosis. Exp Clin Transplant 2020; 18: 206-209.
28. Khattab MA, Zakaria Y, Sadek E, Abd El Fatah AS, Fouad M, Khattab M, et al. Detection of hepatitis C virus (HCV) RNA in the peripheral blood mononuclear cells of HCV-infected patients following sustained virologic response. Clin Exp Med 2023; 23: 131-140.
29. Saad ZM, Ghany WAE, Khalifa R, Higazi A, Al-Shazly M, Said M, et al. Occult HCV infection in liver transplanted patients: frequency and consequences. Clin Exp Hepatol 2022; 8: 125-131.
30. Namayandeh M, Jamalidoust M, Heydari Marandi N, Aliabadi N, Ziyaeyan A, Pouladfar G, et al. Hepatitis C virus genotypes in patients with chronic hepatitis C infection in southern Iran from 2016 to 2019. Microbiol Immunol 2020; 64: 762-767.
31. Keikha M, Eslami M, Yousefi B, Ali-Hassanzadeh M, Kamali A, Yousefi M, et al. HCV genotypes and their determinative role in hepatitis C treatment. Virusdisease 2020; 31: 235-240.
32. Tsai MC, Hung CH, Lu SN, Wang JH, Chen CH, Kee KM, et al. The incidence of resistance-associated variants to NS5A in HCV subtypes 1a and 1b in Taiwan. Biomed J 2021; 44(6 Suppl 1): S126-S131.
33. Ryan P, Valencia J, Cuevas G, Amigot-Sanchez R, Martínez I, Lazarus JV, et al. Decrease in active hepatitis C infection among people who use drugs in Madrid, Spain, 2017 to 2023: a retrospective study. Euro Surveill 2024; 29: 2300712.
34. Scarpetta M, Kanner R, Menezes NP, McDonell CC, Bruneau J, Page K, et al. Getting to full disclosure: HCV testing and status disclosure behaviors among PWID and their injecting partners. BMC Public Health 2025; 25: 1707.
35. Goel A, Rewari BB, Sharma M, Konath NM, Aggarwal R. Seroprevalence and burden of hepatitis C virus infection in WHO South-East Asia Region: A systematic review. J Gastroenterol Hepatol 2022;37:964-972.
36. Torre P, Festa M, Sarcina T, Masarone M, Persico M. Elimination of HCV Infection: Recent Epidemiological Findings, Barriers,and Strategies for the Coming Years. Viruses 2024; 16: 1792.
| Files | ||
| Issue | Vol 18 No 3 (2026) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v18i3.21665 | |
| Keywords | ||
| Hepatitis C Hepatitis C antibodies Hepatitis C antigens Drug users | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Haghi Navand A, Azaran A, Bavi M, Yousefi F, Dastorpour M, Makvandi M. High prevalence of seronegative occult hepatitis C in high-risk individuals. Iran J Microbiol. 2026;18(3):382-392.
