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

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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 1
Externally publishedYes

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Keywords

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

ASJC Scopus subject areas

  • Virology

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