title: Simplified PCR protocols for INNO-LiPA HBV Genotyping and INNO-LiPA HBV PreCore assays.
abstract: BACKGROUND
  INNO-LiPA HBV Genotyping (LiPA HBV GT) and INNO-LiPA HBV PreCore (LiPA HBV PC) are commercially available assays for hepatitis B virus (HBV) characterization. These assays are labor-intensive and may be prone to exogenous DNA contamination due to their use of nested PCR amplification procedures and lack of contamination control measures.
  
  
  OBJECTIVE
  Standardized, single-round INNO-LiPA PCR amplification protocols incorporating uracil N-glycosylase and automated sample processing by the MagNA Pure LC instrument were evaluated.
  
  
  METHODS
  HBV standards containing 10,000, 1000, 100, 10, and 0 IU/mL were analyzed to determine the analytical sensitivity and reproducibility of these modified procedures. One hundred clinical serum specimens with viral titers ranging from 390 to 16,900,000 IU/mL were tested by modified LiPA HBV GT, while 34 specimens with viral titers ranging from 378 to 11,600,000 IU/mL were tested by modified LiPA HBV PC.
  
  
  RESULTS
  Modified LiPA HBV GT and LiPA HBV PC each yielded analytical sensitivities of 100% at an HBV DNA level of 1000 IU/mL and 90% at a level of 100 IU/mL. Among clinical specimens, success rates for both INNO-LiPA procedures were > or =94%.
  
  
  CONCLUSIONS
  Both modified INNO-LiPA procedures were sensitive and reproducible, with improved efficiency and suitability for routine laboratory use.
authors.affiliation: Department of Pathology and Laboratory Medicine, King Faisal Specialist Hospital, Riyadh, Saudi Arabia.
authors.forename: Mohammed O
authors.initials: MO
authors.lastname: Qutub
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Germer
authors.forename: Sjoerd P H
authors.initials: SP
authors.lastname: Rebers
authors.forename: Jayawant N
authors.initials: JN
authors.lastname: Mandrekar
authors.forename: Marcel G H M
authors.initials: MG
authors.lastname: Beld
authors.forename: Joseph D C
authors.initials: JD
authors.lastname: Yao
mesh_terms: D004279:DNA, Viral / Q000032:analysis*; D005838:Genotype; D006509:Hepatitis B / Q000175:diagnosis*; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral