Clinical evaluation of the branched DNA assay for hepatitis B virus DNA detection in patients with chronic hepatitis B lacking hepatitis B e antigen and treated with interferon‐α

Habersetzer, François; Zoulim, Fabien; Jusot, Jean François; Zhang, X.; Trabaud, Mary‐Anne; Chevallier, Philippe; Chevallier, M.; Ahmed, Si Nafa Si; Sepetjan, M.; Comanor, Lorraine; Minor, J.; Trépo, Christian

doi:10.1046/j.1365-2893.1998.00128.x

Cited by 10 publications

(5 citation statements)

References 42 publications

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“…In addition, we observed a strong, significant relationship between HBV DNA levels obtained by the CAP/CTM v2.0 assay and those obtained by the third-generation bDNA-based assay. The use of the third-generation bDNA assay as a comparator was justified by the facts that this assay is accurate, precise, reproducible, and well calibrated to the World Health Organization HBV DNA standard and that it quantifies HBV DNA levels independently of the HBV genotype, due to the presence of a large number of capture and extender probes located at various positions along the HBV genome (7,11). There was also an excellent correspondence between versions 1.0 and 2.0 of the CAP/CTM assay in plasma in our experiments, regardless of the HBV genotype.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2010

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The detection and quantification of hepatitis B virus (HBV) DNA are essential for the diagnosis and treatment of chronic HBV infection. The use of real-time PCR assays for HBV DNA quantification is strongly recommended. The goal of this study was to evaluate the intrinsic characteristics and clinical performance of version 2.0 (v2.0) of the Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) assay, a fully automated platform for HBV DNA quantification in serum or in plasma with a claimed lower limit of detection of 20 IU/ml and a claimed upper limit of quantification of 1.7 ؋ 10 8 IU/ml. The specificity of the assay was 99% (95% confidence interval, 94.7 to 100%). Intra-assay and interassay coefficients of variation ranged from 0.21% to 2.67% and from 0.65% to 2.25%, respectively. The calibration of the assay was found to be satisfactory. Study of blood specimens from patients infected with HBV genotypes A to F showed good correspondence between HBV DNA levels measured by the CAP/CTM v2.0 assay, version 1.0 of the same assay, and the third-generation "branched DNA" assay. The CAP/CTM v2.0 assay quantified HBV DNA levels in serum or plasma from the same patients equally. In conclusion, the new version of the CAP/CTM assay is sensitive, specific, and reproducible. It accurately quantifies HBV DNA levels in patients chronically infected with HBV genotypes A to F. Improvements made to ensure equal quantification of HBV DNA in serum and plasma have been successful. Overall, the CAP/CTM assay, version 2.0, is well suited to monitoring clinical HBV DNA levels according to current clinical practice guidelines.Chronic hepatitis B virus (HBV) infection is associated with a large spectrum of liver diseases, ranging from a low-viremia inactive carrier state to chronic active hepatitis, which may subsequently evolve toward cirrhosis and hepatocellular carcinoma (HCC). Morbidity and mortality are linked to the persistence of viral replication, and hepatic complications develop in 15% to 40% of patients chronically infected with HBV. Overall, HBV-related end-stage liver disease and HCC are responsible for more than 750,000 deaths worldwide per year (5).The detection and quantification of HBV DNA are essential for diagnosing ongoing HBV infection and establishing the prognosis of related liver disease, influence the decision to treat, and are indispensable for monitoring the virological response to antiviral therapy and the emergence of resistance in order to tailor therapy (4). A number of HBV DNA detection and quantification assays are available. For many years, such methods were based on either hybrid capture, signal amplification by means of branched DNA (bDNA) technology, or classical PCR, all of which suffered from poor analytical sensitivity and a narrow range of HBV DNA quantification (3). More recently, assays based on real-time PCR quantification have been developed. Their use for the routine detection and quantification of HBV DNA is recommended, because of their excellent analytical sensitivity (lower limit of detection, 10 to 2...

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Section: Discussionmentioning

confidence: 99%

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

et al. 2010

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“…The pathogenesis of liver damage is immunemediated where the role of cytokines is crucial. Several studies have failed to show correlation between HBV DNA and serum aminotransferases, which is additional evidence that HBV DNA does not correlate with severity of liver disease [13][14] .…”

Section: Discussionmentioning

confidence: 99%

Low Viral Load Does Not Exclude Significant Liver Damage in Patients with Chronic HBV Infection in Bangladesh

Al‐Mahtab

Rahman

Kamal

et al. 2009

Bangabandhu Sheikh Mujib Medical University Journal

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Background: In general, it is assumed that patients with chronic hepatitis B virus (HBV) infection with high viral load exhibit increased liver damages. Accordingly, the treatment guidelines emphasize on reducing viral load in chronic HBV carriers. The ethical and scientific basis of these observations was mainly accumulated from investigations from developed countries of the world. More than 80% chronic HBV carriers live in the developing nations of the world, but little is known about relationship between HBV viral load and extent of liver damages in these countries. In this study, we addressed this issue to provide insights about this. Methods: In this retrospective study we reviewed the records of 210 chronic hepatitis B (CHB) patients from our pool of 561 Bangladeshi CHB patients. All of these 210 patients had low HBV DNA (<10 5 copies/ml by PCR). Of them 16 were HBeAg +ve and rest 194 HBeAg -ve. They have also been tested for other serologic markers of HBV (i.e. HBsAg, anti-HBe), HCV (i.e. anti-HCV) and serum alaninetransaminase (ALT) level. All patients also underwent per-cutaneous liver biopsy. Results: 37.5% (6/16) HBeAg +ve patients with low HBV DNA had significant hepatic necro-inflammation (HAI-NI >7), whereas this figure was 31.44% (61/194) in case of HBeAg -ve patients. On the other hand significant hepatic fibrosis (HAI-F >3) was observed in 31.25% (5/16) and 14. 4% (28/194) in HBeAg +ve and -ve patients respectively. Conclusion: This study shows that a correlation could not be established between viral load and liver damage in patients with CHB in Bangladesh. A significant percentage of patients with low HBV DNA may have marked hepatic necro-inflammation and fibrosis, more so in case of HBeAg +ve CHB. Further study may be needed to find out the influence of other factors on liver damages in CHB patients in developing nations like Bangladesh, where about 8 million chronic HBV carriers are living. Most of these patients have not been characterized and treatment modalities have not been defined for them. Our study may suggest the research direction for management of these cases.

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“…Numerous assays are available for detection of HBV DNA, such as the branched DNA (bDNA) assay (6,13), DNA hybridization assays (9,24), and quantitative PCR (1,12,22). Some of these assays have only limited sensitivity, however, and detection by PCR may be considered to be laborious and susceptible to contamination.…”

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confidence: 99%

Quantitative Detection of Hepatitis B Virus DNA by Real-Time Nucleic Acid Sequence-Based Amplification with Molecular Beacon Detection

Yates¹,

Penning²,

Goudsmit

et al. 2001

J Clin Microbiol

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We have developed a hepatitis B virus (HBV) DNA detection and quantification system based on amplification with nucleic acid sequence-based amplification (NASBA) technology and real-time detection with molecular beacon technology. NASBA is normally applied to amplify single-stranded target RNA, producing RNA amplicons. In this work we show that with modifications like primer design, sample extraction method, and template denaturation, the NASBA technique can be made suitable for DNA target amplification resulting in RNA amplicons. A major advantage of our assay is the one-tube, isothermal nature of the method, which allows high-throughput applications for nucleic acid detection. The homogeneous real-time detection allows a closed-tube format of the assay, avoiding any postamplification handling of amplified material and therefore minimizing the risk of contamination of subsequent reactions. The assay has a detection range of 10 3 to 10 9 HBV DNA copies/ml of plasma or serum (6 logs), with good reproducibility and precision. Compared with other HBV DNA assays, our assay provides good sensitivity, a wide dynamic range, and high-throughput applicability, making it a viable alternative to those based on other amplification or detection methods.Infection with hepatitis B virus (HBV) can cause a spectrum of liver diseases, such as fulminant or chronic hepatitis, cirrhosis, and hepatocellular carcinoma. About 300 million people throughout the world suffer from chronic HBV infection, and 2 billion people have markers indicating past infection with HBV (15,25).HBV is the smallest DNA virus known, and its genome shows a highly compact organization. A unique aspect in the HBV replication cycle is that a pregenomic mRNA serves as a template for the synthesis of the first viral DNA strand by the reverse transcriptase (RT) polymerase of HBV (35). The RNase H activity of the HBV DNA polymerase removes the mRNA during this process, and synthesis of the complementary second DNA strand is then started, generating a partially double-stranded DNA molecule for packaging in virions. When the virus enters the host, this molecule is extended into a fully double-stranded DNA molecule, thus starting a new replication cycle (10,27,28).HBV DNA can be detected in the blood in more than 90% of infected hosts who are positive for hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg). The use of detection and quantification of HBV DNA has become the preferred method for measuring the quantity of infectious particles and provides important diagnostic and prognostic information, mainly as a marker of virus replication (5,14,20,29,30).Numerous assays are available for detection of HBV DNA, such as the branched DNA (bDNA) assay (6, 13), DNA hybridization assays (9, 24), and quantitative PCR (1,12,22). Some of these assays have only limited sensitivity, however, and detection by PCR may be considered to be laborious and susceptible to contamination. Since HBV DNA loads are highly variable and require a broad dynamic detection range ...

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Clinical evaluation of the branched DNA assay for hepatitis B virus DNA detection in patients with chronic hepatitis B lacking hepatitis B e antigen and treated with interferon‐α

Cited by 10 publications

References 42 publications

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

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

Low Viral Load Does Not Exclude Significant Liver Damage in Patients with Chronic HBV Infection in Bangladesh

Quantitative Detection of Hepatitis B Virus DNA by Real-Time Nucleic Acid Sequence-Based Amplification with Molecular Beacon Detection

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