Investigation of putative multisubtype hepatitis C virus infections in vivo by heteroduplex mobility analysis of core/envelope subgenomes.
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.
Li
Hui
H
Department of Laboratory Medicine, University of Washington Medical Center, Seattle, Washington, USA.
Thomassen
Lisa V
LV
Majid
Ayaz
A
McMahon
Brian J
BJ
Bruden
Dana
D
McArdle
Susan
S
Bano
Nazneen
N
Chung
Minjun
M
Carithers
Robert L
RL
Perkins
James D
JD
Sullivan
Daniel G
DG
Gretch
David R
DR
D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D016172:DNA Fingerprinting; D005838:Genotype; D016174:Hepacivirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D019698:Hepatitis C, Chronic / Q000821:virology*; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008099:Liver / Q000821:virology; D008198:Lymph Nodes / Q000821:virology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000235:genetics*; D044967:Serum / Q000821:virology; D014758:Viral Core Proteins / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics
D020121:5' Untranslated Regions; D012367:RNA, Viral; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; C114224:envelope protein E1, GB virus C; C069130:nucleocapsid protein, Hepatitis C virus