Simplified PCR protocols for INNO-LiPA HBV Genotyping and INNO-LiPA HBV PreCore assays

Mohammed O. Qutub, Jeffrey J. Germer, Sjoerd P.H. Rebers, Jayawant N. Mandrekar, Marcel G.H.M. Beld, Joseph D.C. Yao

Research output: Contribution to journalArticle

15 Scopus citations

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. Study design: 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 ≥94%. Conclusions: Both modified INNO-LiPA procedures were sensitive and reproducible, with improved efficiency and suitability for routine laboratory use.

Original languageEnglish (US)
Pages (from-to)218-221
Number of pages4
JournalJournal of Clinical Virology
Volume37
Issue number3
DOIs
StatePublished - Nov 1 2006

Keywords

  • Basal core promoter
  • Genotyping
  • Hepatitis B virus
  • INNO-LiPA
  • MagNA Pure LC
  • Precore

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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