title: Comparative quantitative analysis of hepatitis C mutations at amino acids 70 and 91 in the core region by the Q-Invader assay.
abstract: Hepatitis C virus (HCV) is a major worldwide public health problem, and mutations at amino acids 70 and 91 in the genotype 1b core region predict the effectiveness of combination therapy with peginterferon and ribavirin. An assay based on the Q-Invader technology was developed to determine the relative ratios of the mutant to wild-type virus with high sensitivity. The assay detected a minor type plasmid that constituted only 1% of a mixture of plasmids containing wild-type and mutant sequences. The calculated ratios agreed with those of the template DNA. A total of 123 serum samples of HCV in Japan were examined with the Q-Invader assay. The Q-Invader assay detected all of the mutations that were detected by direct sequencing and even some mutants that direct sequencing could not. PCR with mutant specific primers confirmed those mutations found by the Q-Invader assay and not by direct sequencing. The Q-Invader assay, thus, is a useful tool for detecting mutations at positions 70 and 91 in the HCV-1b core region.
authors.affiliation: BML, Inc., Saitama 350-1101, Japan. tado-k@bml.co.jp
authors.forename: Kenichi
authors.initials: K
authors.lastname: Tadokoro
authors.forename: Mariko
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Suzuki
authors.forename: Chie
authors.initials: C
authors.lastname: Tanaka
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Makoto
authors.initials: M
authors.lastname: Nagano
authors.forename: Toru
authors.initials: T
authors.lastname: Egashira
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Kumada
mesh_terms: D019943:Amino Acid Substitution; D000998:Antiviral Agents / Q000627:therapeutic use; D001483:Base Sequence; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000188:drug therapy / Q000821:virology; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use; D009154:Mutation; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D011092:Polyethylene Glycols / Q000627:therapeutic use; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D011994:Recombinant Proteins / Q000627:therapeutic use; D012254:Ribavirin / Q000627:therapeutic use; D017422:Sequence Analysis, DNA; D014758:Viral Core Proteins / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D016898:Interferon-alpha; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D011092:Polyethylene Glycols; D012254:Ribavirin; C100416:peginterferon alfa-2a