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 language | English |
|---|---|
| Pages (from-to) | 76-84 |
| Number of pages | 9 |
| Journal | Journal of Virological Methods |
| Volume | 149 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2008 Apr 1 |
| Externally published | Yes |
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Keywords
- Chronic hepatitis B
- HBV
- Lamivudine resistance
- Seeplex Lami-DR assay
ASJC Scopus subject areas
- Virology
Cite this
Direct detection of lamivudine-resistant hepatitis B virus mutants by a multiplex PCR using dual-priming oligonucleotide primers. / Kim, Jong Kee; Lee, Hyeon Ji; Lee, Young Jo; Chun, Jong Yoon; Lee, In Kyoung; Lim, Young Suk; Suh, Dong Jin; Ko, Sun-Young; Kim, Myeong Hee; Oh, Heung Bum.
In: Journal of Virological Methods, Vol. 149, No. 1, 01.04.2008, p. 76-84.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Direct detection of lamivudine-resistant hepatitis B virus mutants by a multiplex PCR using dual-priming oligonucleotide primers
AU - Kim, Jong Kee
AU - Lee, Hyeon Ji
AU - Lee, Young Jo
AU - Chun, Jong Yoon
AU - Lee, In Kyoung
AU - Lim, Young Suk
AU - Suh, Dong Jin
AU - Ko, Sun-Young
AU - Kim, Myeong Hee
AU - Oh, Heung Bum
PY - 2008/4/1
Y1 - 2008/4/1
N2 - 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.
AB - 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.
KW - Chronic hepatitis B
KW - HBV
KW - Lamivudine resistance
KW - Seeplex Lami-DR assay
UR - http://www.scopus.com/inward/record.url?scp=40849101514&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=40849101514&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2008.01.003
DO - 10.1016/j.jviromet.2008.01.003
M3 - Article
VL - 149
SP - 76
EP - 84
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -