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Journal of Virology, August 2008, p. 7524-7532, Vol. 82, No. 15
0022-538X/08/$08.00+0     doi:10.1128/JVI.02220-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Investigation of Putative Multisubtype Hepatitis C Virus Infections In Vivo by Heteroduplex Mobility Analysis of Core/Envelope Subgenomes{triangledown}

Hui Li,1 Lisa V. Thomassen,2 Ayaz Majid,5 Brian J. McMahon,6 Dana Bruden,6 Susan McArdle,1 Nazneen Bano,1 Minjun Chung,1 Robert L. Carithers,3 James D. Perkins,4 Daniel G. Sullivan,1 and David R. Gretch1,3*

Departments of Laboratory Medicine,1 Pathology,2 Medicine,3 Surgery, University of Washington Medical Center, Seattle, Washington,4 Department of Medicine, University of Miami, Miami, Florida,5 Arctic Investigators Unit, Centers for Disease Control and Prevention, Anchorage, Alaska6

Received 11 October 2007/ Accepted 30 April 2008

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.

* Corresponding author. Mailing address: 325 Ninth Avenue, Box 359690, Seattle, WA 98104. Phone: (206) 341-5216. Fax: (206) 341-5203. E-mail: gretch{at}u.washington.edu

{triangledown} Published ahead of print on 21 May 2008.

Journal of Virology, August 2008, p. 7524-7532, Vol. 82, No. 15
0022-538X/08/$08.00+0     doi:10.1128/JVI.02220-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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