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Journal of Clinical Microbiology, October 2006, p. 3600-3607, Vol. 44, No. 10
0095-1137/06/$08.00+0     doi:10.1128/JCM.00732-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Expertise of Laboratories in Viral Load Quantification, Genotyping, and Precore Mutant Determination for Hepatitis B Virus in a Multicenter Study

Centre National de Référence pour les Hépatites B et C en Transfusion, Département des Agents Transmissibles par le Sang, Institut National de la Transfusion Sanguine, Paris,¹ Laboratoire de Virologie, Centre Hospitalo-Universitaire Pitié-Salpêtrière, Paris,² Laboratoire de Virologie-Bactériologie, Centre Hospitalo-Universitaire, Limoges,³ Laboratoire de Virologie, Centre Hospitalo-Universitaire, Amiens,⁴ Laboratoire de Virologie, Centre Hospitalo-Universitaire, Nantes,⁵ Laboratoire de Virologie, Centre Hospitalo-Universitaire, Rouen,⁶ Laboratoire Alphabio, Marseille,⁷ Laboratoire de Virologie, Centre Hospitalo-Universitaire, Grenoble,⁸ Laboratoire de Virologie-Bactériologie, Centre Hospitalo-Universitaire, Angers,⁹ Unité de Virologie, Inserm U 481 Centre Abrami, Hôpital Beaujon, Clichy,¹⁰ Laboratoire de Virologie Moléculaire, Etablissement Français du Sang, Bretagne, Brest,¹¹ Laboratoire de Virologie-Bactériologie, Centre Hospitalo-Universitaire Henri-Mondor, Créteil,¹² Laboratoire de Virologie-Bactériologie, Centre Hospitalo-Universitaire, Saint-Etienne,¹³ Laboratoire de Virologie, Hôpital Paul Brousse, Villejuif,¹⁴ Service de Santé Publique, Centre Hospitalo-Universitaire Henri-Mondor, Créteil,¹⁵ Laboratoire de Virologie, Centre Hospitalo-Universitaire Purpan, Toulouse,¹⁶ Laboratoire d'Hématologie, Centre Hospitalo-Universitaire, Amiens,¹⁷ Département des Agents Transmissibles par le Sang, Institut National de la Transfusion Sanguine, Paris, France,¹⁸

Received 6 April 2006/ Returned for modification 16 May 2006/ Accepted 22 June 2006

A national evaluation study was performed in 14 specialized laboratories with the objective of assessing their capacities to provide (i) hepatitis B virus (HBV) viral loads (VL), (ii) HBV genotypes, and(iii) identification of precore/core mutants. The panel consisted of 12 HBV DNA-positive samples with VLs from 2.8 to 9.1 log₁₀ copies/ml, different HBV genotypes (A to F), and 3 mutant and 9 wild-type samples at nucleotide 1896. The coefficients of variation of the mean VLs ranged from 2.4% to 10.4% with the Cobas HBV Monitor assay, from 1.8% to 5.5% with the Cobas TaqMan 48, from 1.5 to 26.2% with RealArt HBV PCR, and from 0 to 7% with branched DNA (bDNA). The Cobas Monitor assay underestimated the VLs of genotype F samples, with differences ranging from 1.4 to 2.4 log₁₀ copies/ml. The accuracies of genotype determinations ranged from 33% to 100%, and those of precore mutant determinations ranged from 25 to 100%. This study showed some drawbacks of two widely used assays: (i) Cobas Monitor has a narrow dynamic range and underestimates genotype F sample VLs and (ii) bDNA shows poor sensitivity and may fail to identify patients with low VLs. With higher performance in terms of analytical sensitivity combined with a larger dynamic range and an ability to quantify the main genotypes equally, real-time PCR methods appear more appropriate for accurate monitoring of HBV DNA quantification. Furthermore, the clinical implications of HBV genotyping and the determination of precore/core mutants need to be clearly stated to justify the standardization of these methods.

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