Presence of different hepatitis C virus genotypes in plasma and peripheral blood mononuclear cell samples of Iranian patients with HIV infection

Abstract

Due to the similar routes of transmission, individuals infected with the human immunodeficiency virus (HIV) may become infected with the hepatitis C virus (HCV) simultaneously. The aim of this study was to investigate the frequency of HCV co-infection in Iranian individuals with HIV infection, and to genotype HCV in plasma and PBMC specimens of these patients. From September 2015 to October 2016, a total of 140 Iranian individuals with HIV infection were enrolled in this cross-sectional study. The RNA from plasma and PBMC specimens was extracted, and genomic HCV-RNA was amplified using RT-nested PCR with primers that target 5'-UTR. The HCV genotyping used the RFLP technique. To confirm HCV genotype, 10 randomly selected HCV-positive samples were also submitted for sequencing. The mean age of patients was 35.7 ± 13.5 years (range: 1-66). Out of 140 patients, 62 (44.3 %) were positive for anti-HCV antibodies; among these, viral genomic RNA was detected in 34 (24.3%) and 39 (27.9%) of the plasma and PBMC specimens, respectively. The HCV genotyping showed a similar pattern of subtypes 1a (44% vs 46.2%), 3a (32.4% vs 33.3%), and 1b (17.6% vs 17.9%) in all sera and PBMC samples. It is noteworthy that the HCV genotypes in plasma and PBMC specimens of 6 HCV co-infected patients were not the same. This study reveals that HIV/HCV co-infection is high in Iranian patients (44.3%), especially in people who have high-risk factors (83.9%). Also, HIV/HCV co-infected individuals may have dissimilar HCV genotypes in their plasma and PBMC specimens.

Detection of HCV genome in peripheral blood mononuclear cells of Iranian seropositive and HCV RNA negative in plasma of patients with beta‐thalassemia major: Occult HCV infection

“…After cDNA synthesis, nested PCR was performed in 25 µL of reaction mixture with 1.2 units EX Taq DNA polymerase (TaKaRa Biotechnology [Dalian] Co, Ltd, Shiga, Japan), 2.5 µL 10× Ex Taq buffer (Mg 2+ free), 2 µL MgCl 2 (25 mM), 2 µL dNTPs mixture (25 mM each) and 2.5 µL cDNA was used as a template of the first stage of PCR, and 1.5 µL of the first round PCR product was used as a template for the second stage of PCR. Temperature profiles that were used in the nested‐PCR assay were the same as previously described in detail …”

“…37 were used in the nested-PCR assay were the same as previously described in detail. 25,38 The second round of PCR products for amplification (629 bp for NS5B and 214 bp for 5′-UTR) were visualized after electrophoresis on agarose gel. The products were purified using a High Pure PCR Product Purification Kit (Roche Diagnostic) according to manufacturer's instructions.…”

“…[16][17][18][19][20] It has been shown in some patients that the HCV genotype in PBMC is different from that in the plasma. [21][22][23][24][25] The genotype of HCV RNA in extrahepatic sites is a major prognostic factor for HCV infection treatment process. 25,26 PBMCs are a favorite site for replication of HCV after treatment at least with interferon treatment.…”

“…[21][22][23][24][25] The genotype of HCV RNA in extrahepatic sites is a major prognostic factor for HCV infection treatment process. 25,26 PBMCs are a favorite site for replication of HCV after treatment at least with interferon treatment. 21,27 It is argued that the main cause of reinfection or relapse of HCV infection after liver transplantation is HCV replication in extrahepatic sites.…”

Prevalence of hepatitis C virus among HIV-infected patients

“…In Iran, the prevalence of HCV in the general population and high-risk populations is 0.3% and 32.1% respectively (10). The HCV genotypes 1a, 3a, 1b are the most dominant in different regions of Iran (7,11). Due to sequence diversity and variation of hepatitis C genome, no vaccine is available against HCV infection.…”