Molecular analysis and ex vivo infectivity of seronegative occult hepatitis C virus: a study in single haemodialysis centre
Background: In occult hepatitis C virus infection (OCI), hepatitis C virus ribonucleic acid (HCV RNA) is detectable in peripheral blood mononuclear cells (PBMCs) but is not evident in serum or plasma. Understanding of OCI in patients with seronegative anti-HCV antibodies is limited. Methods: In...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English English |
| Published: |
Penerbit Universiti Sains Malaysia
2024
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/112008/2/112008_Molecular%20analysis%20and%20ex%20vivo.pdf http://irep.iium.edu.my/112008/3/112008_Molecular%20analysis%20and%20ex%20vivo_Scopus.pdf http://irep.iium.edu.my/112008/ http://www.mjms.usm.my/current.html https://doi.org/10.21315/mjms2024.31.2.4 |
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| Summary: | Background: In occult hepatitis C virus infection (OCI), hepatitis C virus ribonucleic
acid (HCV RNA) is detectable in peripheral blood mononuclear cells (PBMCs) but is not evident
in serum or plasma. Understanding of OCI in patients with seronegative anti-HCV antibodies is
limited.
Methods: In this study, six HCV isolates from haemodialysis (HD) patients with
seronegative OCI were identified by molecular assays and phylogenetic analysis. The virus
infectivity was assessed ex vivo using a primary naïve PBMC culture system. HCV isolates obtained
from the PBMCs of 10 patients with chronic HCV infection (CCI) were characterised concurrently
and used as positive controls in the cell culture.
Results: Sequence analysis of the 5ʹ untranslated region (UTR) and non-structural 5B
(NS5B) region revealed that HCV genotype 3 was the most prevalent virus type in both the OCI
and CCI groups. One of the occult HCV isolates was identified as a mixed type. The mean viral load
(log10 RNA copies/106 cells) in the PBMC samples of the OCI group (M = 3.4, SD = 0.7) was lower
than that of the CCI group (M = 4.6, SD = 1.7). Upon culture, de novo OCI-HCV replicates were
detected in five out of six naïve PBMC cultures. Analysis of the replicates showed a single guanine
addition in the domain III of 5’-UTR but the overall molecular structure was retained.
Conclusion: Seronegative OCI is an active form of infection that replicates at a low level
in PBMCs. Seronegative OCI may share the same route of transmission as CCI. The retained viral
competency may have an implication for its persistence. |
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