Document Detail

Rapid detection of lamivudine-resistant hepatitis B virus mutations by PCR-based methods.
MedLine Citation:
PMID:  16960347     Owner:  NLM     Status:  MEDLINE    
Hepatitis B virus (HBV) is one of the major causes of liver disease worldwide, and chronic HBV infection may progress to cirrhosis and hepatocellular carcinoma. Mutations at the active site of DNA polymerase of HBV, tyrosine-methionine-aspartate-aspartate (YMDD) motif, render infected patients resistant to antiviral drug (Lamivudine) therapy. Hence, sensitive and specific methods aimed at detecting the mutants are essential. The purpose of this study was to develop methods for detecting the mutations at YMDD by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and real-time PCR using locked nucleic acid (LNA)-mediated TaqMan probes. The results obtained by these methods were compared with those examined by conventional direct sequencing on serum samples of 77 patients treated with lamivudine. Our results show that both PCR-RFLP and real-time PCR could detect wild type, YMDD, and its mutants, tyrosine-isoleucine-aspartate-aspartate and tyrosine-valine-aspartate-aspartate. In addition, the mixtures of the wild-type virus and its mutants in the serum sample were detected. Importantly, real-time PCR is less time-consuming, and more sensitive for the detection of mixed populations than PCR-RFLP. The real-time PCR with LNA-mediated TaqMan probes is a sensitive, specific and rapid detection method for mutations at the YMDD motif, which will be essential for monitoring patients undergoing lamivudine antiviral therapy.
Thaweesak Chieochansin; Salin Chutinimitkul; Sunchai Payungporn; Apiradee Theamboonlers; Pisit Tangkijvanich; Piyawat Komolmit; Yong Poovorawan
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Tohoku journal of experimental medicine     Volume:  210     ISSN:  0040-8727     ISO Abbreviation:  Tohoku J. Exp. Med.     Publication Date:  2006 Sep 
Date Detail:
Created Date:  2006-09-08     Completed Date:  2006-10-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0417355     Medline TA:  Tohoku J Exp Med     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  67-78     Citation Subset:  IM    
Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/DQ351989;  DQ351990;  DQ351991;  DQ351992;  DQ351993;  DQ351994;  DQ351995;  DQ351996;  DQ351997;  DQ351998;  DQ351999;  DQ352000;  DQ352001;  DQ352002;  DQ352003;  DQ352004;  DQ352005;  DQ352006;  DQ352007;  DQ352008;  DQ352009;  DQ352010;  DQ352011;  DQ352012;  DQ352013;  DQ352014;  DQ352015;  DQ352016;  DQ352017;  DQ352018;  DQ352019;  DQ352020;  DQ352021;  DQ352022;  DQ352023;  DQ352024;  DQ352025;  DQ352026;  DQ352027;  DQ352028;  DQ352029;  DQ352030;  DQ352031;  DQ352032;  DQ352033;  DQ352034;  DQ352035;  DQ352036;  DQ352037;  DQ352038;  DQ352039
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MeSH Terms
Amino Acid Motifs / genetics
DNA Mutational Analysis / methods*
DNA, Viral / chemistry,  genetics,  isolation & purification
DNA-Directed DNA Polymerase / genetics
Drug Resistance, Viral / genetics
Hepatitis B virus / genetics*
Hepatitis B, Chronic / blood,  virology
Lamivudine / pharmacology
Molecular Sequence Data
Mutation / genetics*
Polymerase Chain Reaction / methods*
Reg. No./Substance:
0/DNA, Viral; 134678-17-4/Lamivudine; EC DNA Polymerase

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