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Virology

Characterization of a New Sensitive PCR Assay for Quantification of Viral DNA Isolated from Patients with Hepatitis B Virus Infections

Vincent Thibault, Christian Pichoud, Carolyn Mullen, James Rhoads, Jane B. Smith, Alain Bitbol, Sven Thamm, Fabien Zoulim
Vincent Thibault
1Laboratoire de Virologie-CERVI, Hôpital de la Pitié-Salpêtrière, 83 Bd. de l'Hôpital, 75651 Paris Cedex 14, France
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  • For correspondence: vincent.thibault@psl.aphp.fr
Christian Pichoud
2INSERM, U871, 69003 Lyon, Université Lyon 1, IFR62 Lyon-Est, 69008 Lyon, and Hospices Civils de Lyon, Hôtel Dieu, Service d'Hépatologie et de Gastroentérologie, 69002 Lyon, France
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Carolyn Mullen
3Abbott Molecular Inc., 1300 East Touhy Ave., Des Plaines, Illinois 60018-3315
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James Rhoads
3Abbott Molecular Inc., 1300 East Touhy Ave., Des Plaines, Illinois 60018-3315
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Jane B. Smith
3Abbott Molecular Inc., 1300 East Touhy Ave., Des Plaines, Illinois 60018-3315
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Alain Bitbol
3Abbott Molecular Inc., 1300 East Touhy Ave., Des Plaines, Illinois 60018-3315
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Sven Thamm
3Abbott Molecular Inc., 1300 East Touhy Ave., Des Plaines, Illinois 60018-3315
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Fabien Zoulim
2INSERM, U871, 69003 Lyon, Université Lyon 1, IFR62 Lyon-Est, 69008 Lyon, and Hospices Civils de Lyon, Hôtel Dieu, Service d'Hépatologie et de Gastroentérologie, 69002 Lyon, France
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DOI: 10.1128/JCM.01180-07
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  • FIG. 1.
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    FIG. 1.

    Correlation between HBV DNA determinations within dynamic ranges using the Abbott RealTime and Versant assays for samples from 59 HBV-infected, HBeAg-negative patients for which a quantitative result was obtained with both assays. The identity line (solid line) as well as the best-fit correlation (broken line) is shown (y = 0.9863x − 0.154, R2 = 0.8646).

  • FIG. 2.
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    FIG. 2.

    Correlation between HBV DNA determinations within dynamic ranges using the RealTime and Versant assays for samples from 59 HBV-infected, HBeAg-negative patients according to surface gene variants (genotypes are indicated).

  • FIG. 3.
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    FIG. 3.

    Correlation between HBV DNA determinations within dynamic ranges using the RealTime and Versant assays for samples from 59 HBV-infected, HBeAg-negative patients according to the number of polymerase gene mutations.

  • FIG. 4.
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    FIG. 4.

    Correlation between HBV DNA determinations using the RealTime and other quantitative PCR assays in HBV DNA samples isolated from patients in cohort B for which a quantitative result was obtained with both assays. (Top) CAM assay (n = 66). (Bottom) CAP-CTM assay (n = 101). The best-fit correlation (solid line) is shown (for the CAM assay, y = 0.8653x + 0.3629 and R2 = 0.7668; for the CAP-CTM assay, y = 0.9586x − 0.0543 and R2 = 0.9276).

Tables

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  • TABLE 1.

    Genotypic characterization of HBeAg-negative HBV strains isolated from 93 samples obtained from cohort A

    HBV strain and relevant genotypeNo. of samples with genotype
    Surface gene variants
        A31
        A/G2
        B9
        C7
        D35
        E6
        F2
        G1
    Polymerase gene variants
        Wild-type34
        L180M1
        M204I14
        M204V1
        L180M + M204I3
        L180M + M204V39
        L180M + M204I + M204V1
  • TABLE 2.

    Comparison of the dynamic ranges of three HBV DNA quantification assaysa

    Test and resultNo. of samples with the following result by Abbott RealTime assay:
    Below detection thresholdQuantifiableAbove quantification limit
    Versant assay (n = 93)
        Below detection threshold130
        Quantifiable0590
        Above quantification limit0300
    Amplicor assay (n = 79)
        Below detection threshold000
        Quantifiable0660
        Above quantification limit0130
    TaqMan assay (n = 106)
        Below detection threshold020
        Quantifiable01010
        Above quantification limit030
    • ↵ a The dynamic range of the Abbott RealTime assay was compared with those of the Versant, Amplicor, and TaqMan assays for detecting HBV DNA in 93 samples isolated from cohort A (Versant) and 106 samples isolated from cohort B (TaqMan). The cohort tested with Amplicor is a subgroup of 79 samples from cohort B.

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Characterization of a New Sensitive PCR Assay for Quantification of Viral DNA Isolated from Patients with Hepatitis B Virus Infections
Vincent Thibault, Christian Pichoud, Carolyn Mullen, James Rhoads, Jane B. Smith, Alain Bitbol, Sven Thamm, Fabien Zoulim
Journal of Clinical Microbiology Dec 2007, 45 (12) 3948-3953; DOI: 10.1128/JCM.01180-07

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Characterization of a New Sensitive PCR Assay for Quantification of Viral DNA Isolated from Patients with Hepatitis B Virus Infections
Vincent Thibault, Christian Pichoud, Carolyn Mullen, James Rhoads, Jane B. Smith, Alain Bitbol, Sven Thamm, Fabien Zoulim
Journal of Clinical Microbiology Dec 2007, 45 (12) 3948-3953; DOI: 10.1128/JCM.01180-07
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KEYWORDS

DNA, Viral
hepatitis B virus
Hepatitis B, Chronic
polymerase chain reaction
viral load

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