title: Rapid detection of the hepatitis B virus YMDD mutant using AllGlo™ probes.
abstract: BACKGROUND
  The early detection of hepatitis B virus (HBV) mutants in clinical samples is important when monitoring chronic HBV patients with lamivudine-resistant mutations during lamivudine therapy.
  
  
  METHODS
  The AllGlo™ probes were designed to distinguish between wild-type (YMDD) and mutant (YVDD and YIDD) strains of HBV. The sensitivity and specificity of the assay were evaluated using a series of diluted mixtures of wild-type and mutant plasmids. This assay was compared with direct sequencing and the mutation-specific primer assay.
  
  
  RESULTS
  Each YMDD, YVDD, and YIDD probe only detected its corresponding plasmid. Moreover, the assay correctly identified negative samples from 40 non-HBV infected patients and 100 healthy controls. The detection limit of this assay was 50 copies/ml for YVDD and YIDD. The assay could detect the mutant strains when they were present at ≥10% within a mixed virus population. The assay was fully concordant with direct sequencing in 34 samples (56.7%) and partially concordant in 26 samples (43.3%), and detected more types of the HBV motif than direct sequencing.
  
  
  CONCLUSIONS
  AllGlo™ probe assay is a novel, sensitive and specific assay to detect lamivudine-related HBV mutants, therefore, may be useful for monitoring chronic HBV patients treated with lamivudine.
authors.affiliation: Department of Center Laboratory, Jinan Infectious Disease Hospital, Shandong University, Jinan, Shandong 250021, PR China.
authors.forename: Zhao-lei
authors.initials: ZL
authors.lastname: Feng
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Yu
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Lu
authors.forename: Da-ying
authors.initials: DY
authors.lastname: Geng
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Shi-jun
authors.initials: SJ
authors.lastname: Chen
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D057230:Limit of Detection; D009154:Mutation / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
chemical_list: D017931:DNA Primers; D015342:DNA Probes