Direct detection of lamivudine-resistant hepatitis B virus mutants by a multiplex PCR using dual-priming oligonucleotide primers

Jong Kee Kim, Hyeon Ji Lee, Young Jo Lee, Jong Yoon Chun, In Kyoung Lee, Young Suk Lim, Dong Jin Suh, Sun Young Ko, Myeong Hee Kim, Heung Bum Oh

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Mutations in the YMDD motif of the hepatitis B virus (HBV) polymerase gene increase lamivudine resistance of HBV, highlighting the clinical importance of accurate and sensitive detection of HBV mutants. Using dual-priming oligonucleotide primer technology, an assay that can detect mutations at codons 180 (L528M) and 204 (YVDD, YIDD, and YSDD) by a single-step multiplex PCR was developed. This Seeplex Lami-DR assay was sufficiently sensitive to detect 103 HBV/ml and was able to detect minor mutants comprising as little as 2% of the viral population. Mutants were detected in 57 of 65 serum samples (88%) from patients with chronic hepatitis B who had been treated with lamivudine (median, 32 months; range, 1-83 months). The agreement with direct sequencing was only 38.5% (25/65). Discrepancies between these methods resulted from detection of additional mutants by the Seeplex Lami-DR assay, as confirmed by a novel verification analysis. This assay is not only highly accurate and sensitive, but is also simple and cost-effective, requiring no expensive probes, laborious sequencing procedures, or digestion with restriction enzymes. Accordingly, the Seeplex HBV Lami-DR assay should be considered as a first-line, cost-effective tool for detecting viral mutations in patients with chronic hepatitis B receiving lamivudine therapy.

Original languageEnglish
Pages (from-to)76-84
Number of pages9
JournalJournal of Virological Methods
Volume149
Issue number1
DOIs
Publication statusPublished - 2008 Apr
Externally publishedYes

Keywords

  • Chronic hepatitis B
  • HBV
  • Lamivudine resistance
  • Seeplex Lami-DR assay

ASJC Scopus subject areas

  • Virology

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