Hepatitis B virus DNA quantitation and detection of core promoter, precore and polymerase mutations in chronic hepatitis B: evaluation and clinical usefulness of three new commercial assays.
BACKGROUND
HBV-DNA quantitation, the HBe antigen status and the appearance of mutations in the core promoter, precore and polymerase regions are important elements in the management of chronic HBV infection.


METHODS
We performed a technical evaluation of 3 new kits, affigene HBV VL, affigene HBV mutant VL and affigene HBV DE/3TC assays (Sangtec Molecular Diagnostics) in comparison with the Amplicor HBV Monitor assay (Manual Test, Roche), direct sequencing and direct sequencing/Inno-LIPA HBV DR (Innogenetics), respectively. We evaluated the clinical application of these tests in the management of patients with chronic (HBeAg positive) hepatitis B. Serial sera of 11 chronic HBeAg positive patients were studied before, during and after lamivudine/interferon treatment.


RESULTS
HBV-DNA quantitation detected with affigene HBV VL showed a high correlation with the Amplicor HBV Monitor test (r=0.85). affigene HBV mutant VL (positions G1764A, G1896A) and affigene HBV DE/3TC (positions rtL180M, rtM204V/I) were able to detect a low presence of mutants in a mixed population (wild type and mutant) compared to direct sequencing and Inno-LIPA HBV DR, which identified only the dominant population.


CONCLUSIONS
These three sensitive assays, performed with the same DNA extraction, give clinicians useful information for the management of chronic hepatitis B and for timing treatment.
Zampino
Rosa
R
Internal Medicine and Hepatology, Second University of Naples, Italy.
Marrone
Aldo
A
Finnström
Niklas
N
Eriksson
Therese
T
Törmänen
Vuokko
V
Hedrum
Andres
A
Attanasio
Vittorio
V
Rosario
Pietro
P
Utili
Riccardo
R
Ruggiero
Giuseppe
G
D000293:Adolescent; D000328:Adult; D000998:Antiviral Agents / Q000627:therapeutic use; D002648:Child; D003199:Computer Systems; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D005260:Female; D005814:Genes, Viral; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000097:blood / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use; D019259:Lamivudine / Q000627:therapeutic use; D008297:Male; D020125:Mutation, Missense / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D018894:Reverse Transcriptase Inhibitors / Q000627:therapeutic use; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D019562:Viral Load
D000998:Antiviral Agents; D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens; D016898:Interferon-alpha; D011933:Reagent Kits, Diagnostic; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine; D004259:DNA-Directed DNA Polymerase