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Sensitive Assay for Quantification of Hepatitis B Virus Mutants by Use of a Minor Groove Binder Probe and Peptide Nucleic Acids

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Title: Sensitive Assay for Quantification of Hepatitis B Virus Mutants by Use of a Minor Groove Binder Probe and Peptide Nucleic Acids
Authors: Hige, Shuhei Browse this author
Yamamoto, Yoichi Browse this author
Yoshida, Shigeru Browse this author
Kobayashi, Tomoe Browse this author
Horimoto, Hiromasa Browse this author
Yamamoto, Keiko Browse this author
Sho, Takuya Browse this author
Natsuizaka, Mitsuteru Browse this author
Nakanishi, Mitsuru Browse this author
Chuma, Makoto Browse this author
Asaka, Masahiro Browse this author
Issue Date: Dec-2010
Publisher: American Society for Microbiology
Journal Title: Journal of Clinical Microbiology
Volume: 48
Issue: 12
Start Page: 4487
End Page: 4494
Publisher DOI: 10.1128/JCM.00731-10
Abstract: Lamivudine is the first nucleoside analogue that was shown to have a potent effect on hepatitis B virus (HBV). However, the emergence of resistant or cross-resistant mutants to nucleos(t)ide analogues remains a serious problem. Several assays for the detection and quantification of antiviral resistant mutants have been reported, but it has been difficult to measure the amounts of mutants accurately, especially when the target strain is minor in the mixed population. It has been shown that accurate measurement of a minor strain is difficult as long as matching reaction with a single probe was included in the assay. We developed a new method for the quantification of lamivudine-resistant strains in a mixed virus population by real-time PCR using minor groove binder probes and peptide nucleic acids, and we achieved a wide and measurable range from 3 to 10 log10 copies/ml and a high sensitivity with a discriminative limit of 0.01% of the predominant strain. The clinical significance of measuring substitutions of not only M204 but also L180 residues of HBV polymerase was demonstrated by this method. This assay increases the versatility of a sensitive method for the quantification of a single nucleotide mutation in a heterogeneous population.
Rights: Copyright © 2010, American Society for Microbiology
Type: article (author version)
Appears in Collections:北海道大学病院 (Hokkaido University Hospital) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 髭 修平

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