Clin Mol Hepatol.  2013 Dec;19(4):399-408. 10.3350/cmh.2013.19.4.399.

Performance evaluation of the HepB Typer-Entecavir kit for detection of entecavir resistance mutations in chronic hepatitis B

• Ahn SH
• Chun JY
• Shin SK
• Park JY
• Yoo W
• Hong SP
• Kim SO
• Han KH

• KMID: 1730886
• DOI: http://doi.org/10.3350/cmh.2013.19.4.399

Abstract

BACKGROUND/AIMS
Molecular diagnostic methods have enabled the rapid diagnosis of drug-resistant mutations in hepatitis B virus (HBV) and have reduced both unnecessary therapeutic interventions and medical costs. In this study we evaluated the analytical and clinical performances of the HepB Typer-Entecavir kit (GeneMatrix, Korea) in detecting entecavir-resistance-associated mutations.
METHODS
The HepB Typer-Entecavir kit was evaluated for its limit of detection, interference, cross-reactivity, and precision using HBV reference standards made by diluting high-titer viral stocks in HBV-negative human serum. The performance of the HepB Typer-Entecavir kit for detecting mutations related to entecavir resistance was compared with direct sequencing for 396 clinical samples from 108 patients.
RESULTS
Using the reference standards, the detection limit of the HepB Typer-Entecavir kit was found to be as low as 500 copies/mL. No cross-reactivity was observed, and elevated levels of various interfering substances did not adversely affect its analytical performance. The precision test conducted by repetitive analysis of 2,400 replicates with reference standards at various concentrations showed 99.9% agreement (2398/2400). The overall concordance rate between the HepB Typer-Entecavir kit and direct sequencing assays in 396 clinical samples was 99.5%.
CONCLUSIONS
The HepB Typer-Entecavir kit showed high reliability and precision, and comparable sensitivity and specificity for detecting mutant virus populations in reference and clinical samples in comparison with direct sequencing. Therefore, this assay would be clinically useful in the diagnosis of entecavir-resistance-associated mutations in chronic hepatitis B.

Keyword

HBV; Drug; Resistance mutation; Entecavir; MALDI-TOF

MeSH Terms

Adult
Antiviral Agents/*therapeutic use
Cross Reactions
DNA, Viral/blood/standards
Drug Resistance, Viral/*genetics
Genotype
Guanine/*analogs & derivatives/therapeutic use
Hepatitis B virus/genetics
Hepatitis B, Chronic/*drug therapy/genetics
Humans
Mutation
Polymerase Chain Reaction/standards
Reagent Kits, Diagnostic/*standards
Reference Standards
Sequence Analysis, DNA
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/standards
Antiviral Agents
DNA, Viral
Guanine
Reagent Kits, Diagnostic