Peptide nucleic acid array for detection of point mutations in hepatitis B virus associated with antiviral resistance.
The detection of antiviral-resistant hepatitis B virus (HBV) mutations is important for monitoring the response to treatment and for effective treatment decisions. We have developed an array using peptide nucleic acid (PNA) probes to detect point mutations in HBV associated with antiviral resistance. PNA probes were designed to detect mutations associated with resistance to lamivudine, adefovir, and entecavir. The PNA array assay was sensitive enough to detect 10(2) copies/ml. The PNA array assay was able to detect mutants present in more than 5% of the virus population when the total HBV DNA concentration was greater than 10(4) copies/ml. We analyzed a total of 68 clinical samples by this assay and validated its usefulness by comparing results to those of the sequencing method. The PNA array correctly identified viral mutants and has high concordance (98.3%) with direct sequencing in detecting antiviral-resistant mutations. Our results showed that the PNA array is a rapid, sensitive, and easily applicable assay for the detection of antiviral-resistant mutation in HBV. Thus, the PNA array is a useful and powerful diagnostic tool for the detection of point mutations or polymorphisms.
Jang
Hyunjung
H
Panagene Inc., 816 Tamnip-Dong, Youseong-Gu, Daejeon, 305-510, South Korea.
Kim
Jihyun
J
Choi
Jae-jin
JJ
Son
Yeojin
Y
Park
Heekyung
H
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D004279:DNA, Viral / Q000235:genetics; D024882:Drug Resistance, Viral; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D006509:Hepatitis B / Q000821:virology; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology; D008826:Microbial Sensitivity Tests / Q000379:methods; D020125:Mutation, Missense; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D063065:Organophosphonates / Q000494:pharmacology; D020135:Peptide Nucleic Acids; D017354:Point Mutation; D012680:Sensitivity and Specificity
D000998:Antiviral Agents; D004279:DNA, Viral; D063065:Organophosphonates; D020135:Peptide Nucleic Acids; D019259:Lamivudine; C413685:entecavir; D006147:Guanine; C053001:adefovir; D000225:Adenine