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. 2007 Jul;81(14):7710-5.
doi: 10.1128/JVI.02750-06. Epub 2007 May 2.

Ultracentrifugation of serum samples allows detection of hepatitis C virus RNA in patients with occult hepatitis C

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Ultracentrifugation of serum samples allows detection of hepatitis C virus RNA in patients with occult hepatitis C

Javier Bartolomé et al. J Virol. 2007 Jul.

Abstract

Occult hepatitis C virus (HCV) infection of patients with abnormal liver function tests of unknown origin who are anti-HCV and serum HCV RNA negative but who have HCV RNA in the liver has been described. As HCV replicates in the liver cells of these patients, it could be that the amount of circulating viral particles is under the detection limit of the most sensitive techniques. To prove this hypothesis, serum samples from 106 patients with occult HCV infection were analyzed. Two milliliters of serum was ultracentrifuged over a 10% sucrose cushion for 17 h at 100,000 x g(av), where av means average, and HCV RNA detection was performed by strand-specific real-time PCR. Out of the 106 patients, 62 (58.5%) had detectable serum HCV RNA levels after ultracentrifugation, with a median load of 70.5 copies/ml (range, 18 to 192). Iodixanol density gradient studies revealed that HCV RNA was positive at densities of 1.03 to 1.04 and from 1.08 to 1.19 g/ml, which were very similar to those found in the sera of patients with classical chronic HCV infection. Antigenomic HCV RNA was found in the livers of 56 of 62 (90.3%) patients with detectable serum HCV RNA levels after ultracentrifugation, compared to 27 of 44 (61.4%) negative patients (P < 0.001). No differences in the median loads of antigenomic HCV RNA between patients with an those without serum HCV RNA (4.5 x 10(4) [range, 7.9 x 10(2) to 1.0 x 10(6)] versus 2.3 x 10(4) [range, 4.0 x 10(2) to 2.2 x 10(5)]) were found. Alanine aminotransferase and gamma-glutamyl transpeptidase levels, liver necroinflammatory activity, and fibrosis did not differ between both groups. In conclusion, HCV RNA can be detected in the sera of patients with occult HCV infection after circulating viral particles are concentrated by ultracentrifugation.

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Figures

FIG. 1.
FIG. 1.
Neighbor-joining tree constructed with the HCV core nucleotide sequences of the clones of 10 serum HCV RNA-positive patients (GenBank accession numbers DQ839181 to DQ839220) from whom enough serum sample was available to repeat ultracentrifugation and those corresponding to the different HCV genotypes. Bootstrap values ≥70, obtained after 1,000 replicates of the data sheet, are shown in the nodes of the tree.
FIG. 2.
FIG. 2.
HCV RNA detection in fractions obtained from the iodixanol gradients in the serum of a patient with an occult HCV infection (a) or with a classical chronic HCV infection (b).

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References

    1. Bradley, D., K. McCaustland, K. Krawczynski, J. Spelbring, C. Humphrey, and E. H. Cook. 1991. Hepatitis C virus: buoyant density of the factor VIII-derived isolate in sucrose. J. Med. Virol. 34:206-208. - PubMed
    1. Bullock, G. C., D. E. Bruns, and D. M. Haverstick. 2002. Hepatitis C genotype determination by melting curve analysis with a single set of fluorescence resonance energy transfer probes. Clin. Chem. 48:2147-2154. - PubMed
    1. Castillo, I., M. Pardo, J. Bartolome, N. Ortiz-Movilla, E. Rodriguez-Iñigo, S. de Lucas, C. Salas, J. A. Jimenez-Heffernan, A. Perez-Mota, J. Graus, J. M. López-Alcorocho, and V. Carreño. 2004. Occult hepatitis C virus infection in patients in whom the etiology of persistently abnormal results of liver-function tests is unknown. J. Infect. Dis. 189:7-14. - PubMed
    1. Castillo, I., E. Rodriguez-Iñigo, J. Bartolomé, S. de Lucas, N. Ortiz-Movilla, J. M. Lopez-Alcorocho, M. Pardo, and V. Carreño. 2005. Hepatitis C virus replicates in peripheral blood mononuclear cells of patients with occult hepatitis C virus infection. Gut 54:682-685. - PMC - PubMed
    1. Castillo, I., E. Rodríguez-Iñigo, J. M. López-Alcorocho, M. Pardo, J. Bartolomé, and V. Carreño. 2006. Hepatitis C virus replicates in the liver of patients who have a sustained response to antiviral treatment. Clin. Infect. Dis. 43:1277-1283. - PubMed

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