Performance of Version 2.0 of the Cobas AmpliPrep/Cobas TaqMan Real-Time PCR Assay for Hepatitis B Virus DNA Quantification

Chevaliez, Stéphane; Bouvier‐Alias, Magali; Laperche, Syria; Hézode, Christophe; Pawlotsky, Jean‐Michel

doi:10.1128/jcm.01306-10

Cited by 60 publications

(47 citation statements)

References 12 publications

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“…This study is the first to simultaneously compare the performance of these three assays and corroborates prior reports of the performance of CAP/CTM (3,4,6), HP (3,11), and TNAI (10). Sensitivities were similar, with HP and TNAI slightly more sensitive than CAP/CTM in the study, and the sensitivity of the CAP/ CTM assay closely matched previously reported results (6).…”

Section: Discussionsupporting

confidence: 85%

Comparison of Three Roche Hepatitis B Virus Viral Load Assay Formats

et al. 2012

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, were compared to a modified (not FDA-approved) version of the HP assay by automating the DNA extraction using the Total Nucleic Acid Isolation (TNAI) kit on the Cobas AmpliPrep. On average, CAP/CTM measurements were 0.08 log IU/ml higher than HP results (n ‫؍‬ 206), and TNAI results were 0.17 log IU/ml higher than HP results (n ‫؍‬ 166). The limit of detection (LOD), as determined by probit analysis using dilutions of the 2nd HBV international standard, was 10.2 IU/ml for CAP/CTM. The data sets for HP and TNAI were insufficient for probit analysis; however, there was 100% detection at >5 or >10 IU/ml for TNAI and HP, respectively. Linearity was demonstrated between 60 and 2,000,000 IU/ml, with slopes between 0.95 and 0.99 and R 2 values of >0.99 for all assays. Total precision (log percent coefficient of variance [CV]) was between 0.8% and 2.1% at 4.3 log IU/ml and between 1.4% and 4.9% at 2.3 log IU/ml. Correlation of samples, reproducibility, linearity, and LOD were acceptable and similar in all assays. The CAP/CTM assay and, to a lesser extent, the TNAI assay reduced hands-on time due to automation. There were no instances of contamination detected in negative samples during the course of the study, despite testing several samples up to 9.6 log IU/ml. The incidence of false-positive negative controls in HP and CAP/CTM clinical testing was <0.5% over 6 to 7 months of testing. Hepatitis B virus (HBV) is a DNA virus that infects up to 400 million people worldwide and causes up to 5,500 deaths annually in the United States from the resulting liver failure, cirrhosis, and hepatocellular carcinoma (5). In the last decade, significant progress has been made in HBV treatment, with the development of new therapeutics (2,5,8,12) with improved genetic barriers and potency. In addition, the techniques for measuring HBV DNA levels in serum and plasma have improved. HBV DNA levels are used to predict response to therapy, to determine therapy initiation, to monitor resistance to therapy, and to establish treatment success (2,5,8,12). Since HBV DNA levels can vary from very low levels to more than 9 log IU/ml, the most recently approved assays that use real-time PCR to generate results over a large dynamic range are preferred. Other available quantitative methods use signal amplification or conventional PCR. The real-time PCR assays typically incorporate robotic automation, making them attractive for high-throughput laboratories.Roche has released two FDA-approved (in vitro diagnostic [IVD]) HBV real-time PCR viral load assays. The first assay approved was the Cobas TaqMan HBV Test for use with the High Pure System (HP) (11). HP uses a manual 12-well silica-based plate format for DNA extraction, followed by manual inoculation into PCR tubes, which are thermocycled and detected using the TaqMan 48 instrument. Most recently, the Cobas AmpliPrep/Cobas TaqMan HBV Test v2.0 (CAP/CTM) (6) integrates automated sample extraction and PCR assembly using the AmpliPrep instrument with the TaqMan instrument for thermocycling and detect...

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Section: Discussionsupporting

confidence: 85%

Comparison of Three Roche Hepatitis B Virus Viral Load Assay Formats

et al. 2012

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“…Serum levels of ALT, creatinine, phosphorus, HBV DNA, HBeAg, and anti-hepatitis B e antibody were assessed for all patients every 2 to 3 months. Serum HBV DNA levels were measured at baseline and each follow-up visit using either Roche COBAS TaqMan (lower limit of detection, 20 IU/ml; Roche Molecular Systems, Branchburg, NJ) or Abbott m2000 (lower limit of detection, 15 IU/ml; Abbott Diagnostics, Chicago, IL) (20). At baseline, testing for genotypic resistance, which was defined as the detection of HBV variants with substitutions conferring antiviral drug resistance, was performed for all study patients and was repeated for patients who experienced virologic breakthrough during the treatment period.…”

Section: Methodsmentioning

confidence: 99%

Tenofovir Monotherapy versus Tenofovir plus Lamivudine or Telbivudine Combination Therapy in Treatment of Lamivudine-Resistant Chronic Hepatitis B

Lee

Jung

Kim

et al. 2015

Antimicrob Agents Chemother

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d Tenofovir disoproxil fumarate (TDF) monotherapy is a therapeutic option for chronic hepatitis B (CHB) patients infected with hepatitis B virus (HBV) variants resistant to lamivudine (LAM). We evaluated the antiviral efficacy and safety of TDF alone compared to those of TDF plus LAM or telbivudine (LdT) combination in patients harboring HBV variants with genotypic resistance to LAM. This multicenter retrospective study included consecutive patients who had LAM-resistant HBV variants and were treated with TDF alone (monotherapy group) or TDF combined with LAM or LdT (combination therapy group) for at least 6 months. Inverse probability of treatment weighting (IPTW) for the entire cohort was applied to control for treatment selection bias. Overall, 153 patients (33 in the monotherapy group and 120 in the combination therapy group) were analyzed. The overall probability of achieving complete virologic suppression at month 12 was 91.6%: 88.6% in the monotherapy group and 92.6% in the combination therapy group. Combination therapy was not superior to monotherapy in viral suppression before and after IPTW (P ‫؍‬ 0.562 and P ‫؍‬ 0.194, respectively). Hepatitis B e antigen (HBeAg) loss, biochemical response, and virologic breakthrough did not differ between treatment groups. The probabilities of complete virologic suppression were comparable between treatment groups in the subsets according to HBeAg status and HBV DNA levels at baseline. No patient experienced any significant renal dysfunction during the treatment period. In conclusion, TDF monotherapy has antiviral efficacy comparable to that of TDF plus LAM or LdT combination therapy, with a favorable safety profile in CHB patients with LAM-resistant HBV variants. L amivudine (LAM), which is the first oral antiviral drug approved for chronic hepatitis B (CHB) treatment, has been widely prescribed for patients with chronic hepatitis B virus (HBV) infection (1). However, the major drawback of LAM is a low barrier to resistance; resistance to LAM occurs in approximately 24% of patients after 1 year of treatment and progressively increases, such that 70% of patients demonstrate resistance after 5 years of LAM therapy (2, 3).Based on clinical evidence (4, 5), international guidelines recommend adding on adefovir (ADV) therapy rather than switching to ADV or entecavir (ETV) monotherapy as a treatment for CHB patients infected with HBV variants resistant to LAM (6, 7). However, because combination therapy with LAM and ADV has limited antiviral efficacy in LAM-resistant patients, a substantial number of patients show persistent viremia while on the rescue therapy with LAM plus ADV combination, which may then result in selection for multidrug-resistant HBV variants and progression of liver disease (8, 9). Other treatment options for patients harboring LAM-resistant HBV include tenofovir disoproxil fumarate (TDF) with or without other nucleoside analogues. Preceding in vitro and clinical studies demonstrated that TDF has antiviral activity against LAM-resistant HBV isolates as...

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“…Laboratory measurements were assessed for all patients, including serum levels of ALT, creatinine, and HBV DNA and hepatitis B e antigen (HBeAg) and antibody (anti-HBe Ab) every 2 to 3 months. At baseline and at each follow-up visit, serum HBV DNA levels were quantified using the Cobas AmpliPrep/Cobas TaqMan version 2.0 assay (Roche Molecular System, Branchburg, NJ), which has a dynamic range of quantification of 20 to 1.7 ϫ 10 8 IU/ml (1.3 to 8 log 10 IU/ml) (16). HBeAg and anti-HBe Ab were determined using a radioimmunoassay (RIA Elisa Rapid kit; Shin Jin Medics, Seoul, Republic of Korea).…”

Section: Methodsmentioning

confidence: 99%

Efficacy of Entecavir-Tenofovir Combination Therapy for Chronic Hepatitis B Patients with Multidrug-Resistant Strains

Lee

et al. 2014

Antimicrob Agents Chemother

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bThe emergence of multidrug-resistant (MDR) strains of hepatitis B virus (HBV) is a major concern. This study aimed to investigate the efficacy and safety of combination therapy with entecavir (ETV) plus tenofovir disoproxil fumarate (TDF) against MDR HBV. To adjust for differences in baseline characteristics, inverse probability weighting (IPW) using propensity scores for the entire cohort and weighted Cox proportional hazards models were applied. Ninety-three consecutive patients who were treated with ETV-TDF combination therapy for >6 months were included; at baseline, 45 were infected with HBV strains with genotypic resistance to lamivudine (LAM) and ETV (the LAM/ETV-R group), 28 with strains resistant to LAM and adefovir (ADV) (the LAM/ADV-R group), and 20 with strains resistant to LAM, ETV, and ADV (the LAM/ETV/ADV-R group). The median duration of rescue therapy was 13.0 (range, 6.7 to 31.7) months. Seventy-four of 93 patients (79.6%) achieved complete virologic suppression, after a median of 4.5 (95% confidence interval, 3.0 to 6.0) months. The cumulative probability of complete virologic suppression at month 6 was 63.6% (55.7%, 75.0%, and 65.0% in the LAM/ETV-R, LAM/ADV-R, and LAM/ETV/ADV-R groups, respectively). During the treatment period, these probabilities were not significantly different across the resistance profiles before and after IPW (P ‫؍‬ 0.072 and P ‫؍‬ 0.510, respectively). In multivariate analysis, a lower baseline HBV DNA level, but not resistance profiles, was an independent predictor of complete virologic suppression. Renal dysfunction was not observed during the treatment period. In conclusion, rescue therapy with ETV-TDF combination is efficient and safe in patients infected with MDR HBV strains regardless of the antiviral drug resistance profiles.T he goal of chronic hepatitis B (CHB) treatment is to achieve early and sustained suppression of hepatitis B virus (HBV) replication, which is demonstrated to prevent progression of liver disease to cirrhosis and development of hepatocellular carcinoma (1, 2). The availability of nucleos(t)ide analogues such as tenofovir disoproxil fumarate (TDF) and entecavir (ETV), which are more potent than other antiviral drugs, has significantly improved treatment of CHB (3-5). However, many patients were treated with less potent antiviral drugs (i.e., lamivudine [LAM], telbivudine [LdT], and adefovir [ADV]) as first-line therapy and then with sequential monotherapies, which contributed to the development of multidrug resistance (MDR) (6, 7). The emergence of drug-resistant viral strains results in increased viral loads, followed by increases in serum alanine aminotransferase (ALT) levels and subsequent progression of liver disease (8-10). In the absence of treatment intervention with appropriate rescue therapy based on cross-resistance profiles, patients are at significant risk of hepatic decompensation (11). Previous antiviral treatment history may impair the antiviral efficacy of rescue therapy to induce viral suppression; therefore, the choice of ...

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Performance of Version 2.0 of the Cobas AmpliPrep/Cobas TaqMan Real-Time PCR Assay for Hepatitis B Virus DNA Quantification

Cited by 60 publications

References 12 publications

Comparison of Three Roche Hepatitis B Virus Viral Load Assay Formats

Comparison of Three Roche Hepatitis B Virus Viral Load Assay Formats

Tenofovir Monotherapy versus Tenofovir plus Lamivudine or Telbivudine Combination Therapy in Treatment of Lamivudine-Resistant Chronic Hepatitis B

Efficacy of Entecavir-Tenofovir Combination Therapy for Chronic Hepatitis B Patients with Multidrug-Resistant Strains

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