Research Article
Investigation of Putative Multisubtype Hepatitis C Virus Infections In Vivo by Heteroduplex Mobility Analysis of Core/Envelope Subgenomes

Hui Li, Lisa V. Thomassen, Ayaz Majid, Brian J. McMahon, Dana Bruden, Susan McArdle, Nazneen Bano, Minjun Chung, Robert L. Carithers, James D. Perkins, Daniel G. Sullivan and David R. Gretch

Journal of Virology, 2008, 82(15), 7524-7532. DOI: 10.1128/JVI.02220-07

Abstract

The frequency that multiple different subtypes of hepatitis C virus (HCV) simultaneously infect a given individual is controversial. To address this question, heteroduplex mobility analysis (HMA) of portions of the HCV core and envelope 1 region was optimized for sensitive and specific detection of mixtures of HCV genomes of different genotype or subtype. Using the standard HCV genotyping approach of 5`-untranslated region (UTR) analysis, 28 of 374 (7.5%) chronic hepatitis C research subjects were classified as having either multiple-subtype HCV infections (n = 21) or switching HCV subtypes over time (n = 7), the latter pattern implying viral superinfection. Upon retesting of specimens by HMA, 25 of 28 multiple-subtype results could not be reproduced. All three patients with positive results were injection drug users with potential multiple HCV exposures. To address the hypothesis of tissue sequestration of multiple-subtype HCV infections, liver (n = 22), peripheral blood mononuclear cell (n = 13), perihepatic lymph node (n = 16), and serum (n = 19) specimens from 23 subjects with end-stage hepatitis C were collected and analyzed by the HMA technique. Whereas 5`-UTR results implicated mixed-subtype HCV infections in 2 subjects, HMA testing revealed no evidence of a second HCV subtype in any tissue compartment (0 of 70 compartments [0%]) or within any given subject (0 of 23 subjects [0%]). In summary, a large proportion of mixed-genotype and switching-genotype patterns generated by 5`-UTR analysis were not reproducible using the HMA approach, emphasizing the need for additional study.

ASCI-ID: 205-711

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