scholarly journals The New Aptima HBV Quant Real-Time TMA Assay Accurately Quantifies Hepatitis B Virus DNA from Genotypes A to F

2017 ◽  
Vol 55 (4) ◽  
pp. 1211-1219 ◽  
Author(s):  
Stéphane Chevaliez ◽  
Claude Dauvillier ◽  
Fabienne Dubernet ◽  
Jean-Dominique Poveda ◽  
Syria Laperche ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Limit Of Detection ◽  
Antiviral Treatment ◽  
Hbv Dna ◽  
Hbv Genotypes ◽  
B Virus ◽  
Test Version ◽  
Related Liver Disease

ABSTRACT Sensitive and accurate hepatitis B virus (HBV) DNA detection and quantification are essential to diagnose HBV infection, establish the prognosis of HBV-related liver disease, and guide the decision to treat and monitor the virological response to antiviral treatment and the emergence of resistance. Currently available HBV DNA platforms and assays are generally designed for batching multiple specimens within an individual run and require at least one full day of work to complete the analyses. The aim of this study was to evaluate the ability of the newly developed, fully automated, one-step Aptima HBV Quant assay to accurately detect and quantify HBV DNA in a large series of patients infected with different HBV genotypes. The limit of detection of the assay was estimated to be 4.5 IU/ml. The specificity of the assay was 100%. Intra-assay and interassay coefficients of variation ranged from 0.29% to 5.07% and 4.90% to 6.85%, respectively. HBV DNA levels from patients infected with HBV genotypes A to F measured with the Aptima HBV Quant assay strongly correlated with those measured by two commercial real-time PCR comparators (Cobas AmpliPrep/Cobas TaqMan HBV test, version 2.0, and Abbott RealTi m e HBV test). In conclusion, the Aptima HBV Quant assay is sensitive, specific, and reproducible and accurately quantifies HBV DNA in plasma samples from patients with chronic HBV infections of all genotypes, including patients on antiviral treatment with nucleoside or nucleotide analogues. The Aptima HBV Quant assay can thus confidently be used to detect and quantify HBV DNA in both clinical trials with new anti-HBV drugs and clinical practice.

scholarly journals Diagnosis and Monitoring of Hepatitis B Virus Infection Using the Cobas® HBV Test for Use on the Cobas® 4800 System

2021 ◽  
Vol 9 (3) ◽  
pp. 573
Author(s):  
Valérie Ortonne ◽  
Mélanie Wlassow ◽  
Magali Bouvier-Alias ◽  
Giovana Melica ◽  
Jean-Dominique Poveda ◽  
...  
Keyword(s):  
Clinical Practice ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Limit Of Detection ◽  
Clinical Performance ◽  
Hbv Dna ◽  
Nucleic Acid Amplification ◽  
Hbv Genotypes ◽  
B Virus ◽  
System 2

(1) Background: Sensitive and accurate nucleic acid amplification technologies are now recommended for hepatitis B virus (HBV) DNA detection and quantification in clinical practice to diagnose and monitor hepatitis B infection. The aim of this study was to assess the analytical and clinical performance of the cobas® HBV Test on the cobas® 4800 System. (2) Methods: Standard panel and clinical specimens were tested in parallel with three different real-time commercial PCR assays including the cobas ® HBV Test, the Cobas® AmpliPrep/Cobas® TaqMan HBV Test v2.0 and Alinity™ m HBV assay. (3) Results: The specificity of the cobas® HBV Test was 97.9%. The limit of detection was estimated to be 2.1 IU/mL. Intra-assay and interassay coefficients of variation varied from 0.14% to 1.92% and 2.16% to 12.02%, respectively. HBV DNA levels in patients infected with different HBV genotypes strongly correlated with those measured by the two other commercial comparators assays. (4) Conclusions: The cobas® HBV Test can be confidently used to detect and accurately quantify HBV DNA in clinical practice as well as in clinical trials with the new anti-HBV drugs currently in development.

Hepatitis B virus (HBV) DNA quantification and HBV genotypes detection in dried blood specimens

2003 ◽  
Vol 38 ◽  
pp. 112
Author(s):  
R. Jardi ◽  
F. Rodriguez-Frias ◽  
M. Buti ◽  
X. Costa ◽  
A. Valdes ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Dna Quantification ◽  
Hbv Genotypes ◽  
B Virus ◽  
Blood Specimens

Real-time quantitation of hepatitis B virus (HBV) DNA in tumorous and surrounding tissue from patients with hepatocellular carcinoma

2002 ◽  
Vol 68 (4) ◽  
pp. 494-499 ◽  
Author(s):  
Isabella Zanella ◽  
Angelo Rossini ◽  
Daniela Domenighini ◽  
Alberto Albertini ◽  
Elisabetta Cariani
Keyword(s):  
Hepatocellular Carcinoma ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Hbv Dna ◽  
Surrounding Tissue ◽  
B Virus

scholarly journals Real Time PCR HBV-DNA Analysis in HBsAg Positive Patients

2015 ◽  
Vol 10 (2) ◽  
pp. 95-99
Author(s):  
Wasila Rahman ◽  
Muhammad Rabiul Hossain ◽  
Arif Ahmed Khan ◽  
Debashish Saha ◽  
SM Mahbubul Alam ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Antiviral Therapy ◽  
Real Time ◽  
Real Time Pcr ◽  
Armed Forces ◽  
Hbv Dna ◽  
Health Concern ◽  
B Virus ◽  
Pcr Method

Introduction: The hepatitis B virus is a global public health concern and leading cause of chronic liver disease in Bangladesh. For the diagnosis and monitoring of treatment of Hepatitis B virus infection, HBV-DNA detection and quantification is now extensively used worldwide.Objectives: The objective of this study was to detect HBV-DNA by real time PCR method in HBsAg positive patients, to compare the results of HBVDNA detection with HBeAg and Anti-HBe and to monitor the response after antiviral therapy in chronic hepatitis B patients and also to observe the intensity of hepatitis B infection in relation to age and sex.Methods: This was a cross sectional type of study conducted in Armed Forces Institute of Pathology (AFIP), Dhaka Cantonment. In this study, 56 sera of HBsAg positive patients were selected who all were subjected to do HBV-DNA (real time PCR) analysis during the period of 29 July to 30 0ctober, 2013.Results: Out of 56 HBsAg positive patients, HBV-DNA was detected in 34 patients. Among these, 8 (23.5%) patients were HBeAg positive, 16 (47%) patients were anti-HBe positive and 10 (29.5%) were negative for both HBeAg and anti-HBe. Age limit of patients was up to 60 years. HBV-DNA positive patients showed male predominance; 26 (76.5%) patients were male and 8 (23.5%) patients were female. Mean age of the patients was 35±14 years. Among 56 HBsAg positive patients, fifteen were receiving antiviral therapy. Out of them, HBV-DNA was decreased among 4 patients and could not be detected among 11 patients.Conclusion: Real time PCR method of detection of HBV-DNA is very important in patients who are HBeAg negative and this method is also applied to monitor treatment response to antivirals and to detect occult HBV infections immune control phase and also to detect reactivation of HBV cases.Journal of Armed Forces Medical College Bangladesh Vol.10(2) 2014

scholarly journals Automated Quantitative Analysis of Hepatitis B Virus DNA by Using the Cobas Amplicor HBV Monitor Test

1999 ◽  
Vol 37 (9) ◽  
pp. 2793-2797 ◽  
Author(s):  
Ulrika Noborg ◽  
Annkatrin Gusdal ◽  
Eva K. Pisa ◽  
Anders Hedrum ◽  
Magnus Lindh
Keyword(s):  
Quantitative Analysis ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Antiviral Treatment ◽  
Colorimetric Detection ◽  
Hbv Dna ◽  
Linear Range ◽  
Virus Concentration ◽  
B Virus ◽  
The Mean

A highly sensitive method of quantitative analysis of hepatitis B virus (HBV) DNA in serum, the Cobas Amplicor HBV Monitor (Cobas-AM) test, was evaluated. Following a manual extraction of viral DNA, amplification, colorimetric detection, and quantitative determination are all automatically performed in the Cobas analyzer. Serially diluted samples with known HBV DNA concentrations were analyzed blindly. All samples with a virus concentration of 400 copies/ml and 83% of samples with a virus concentration of 100 copies/ml could be detected. A linear correlation between input HBV DNA and measured HBV DNA was seen in the range from 100 to 105 copies/ml. The mean coefficient of variation was 29.6% for all input levels and 18.9% for HBV DNA concentrations above 400 copies/ml. Samples with an HBV DNA level above 109 copies/ml could be reproducibly measured after predilution to 10−4 or 10−6 in negative serum; however, the level was underestimated if target DNA after dilution was still above the linear range of the assay. Quantitative results of the Cobas-AM test were interchangeable with measurements by the manual microwell plate version of Amplicor HBV Monitor (MWP-AM); the mean ratio for log Cobas-AM results/log MWP-AM results was 0.97 (standard error of the mean, 0.007) when serum samples from 153 chronic carriers were analyzed. The test should be of value for clinical assessment of chronic carriers and for monitoring the response to antiviral treatment. A limitation is the relatively narrow linear range of the assay, requiring predilution of high-titer (mainly hepatitis B e-antigen-positive) samples.

scholarly journals Rebound of Hepatitis B Virus Replication in HepG2 Cells after Cessation of Antiviral Treatment

2002 ◽  
Vol 76 (16) ◽  
pp. 8148-8160 ◽  
Author(s):  
Ayman M. Abdelhamed ◽  
Colleen M. Kelley ◽  
Thomas G. Miller ◽  
Phillip A. Furman ◽  
Harriet C. Isom
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Drug Treatment ◽  
Time Course ◽  
Antiviral Treatment ◽  
Recombinant Baculovirus ◽  
Antiviral Agent ◽  
Hbv Dna ◽  
Hbv Replication ◽  
B Virus

ABSTRACT Treatment of patients with lamivudine (3TC) results in loss of detectable levels of hepatitis B virus (HBV) DNA from serum; however, the relapse rate, with regard to both reappearance of virus in the bloodstream and hepatic inflammation, is high when therapy is terminated. Although the rebound observed in patients has also been seen in animal hepadnavirus models, rebound has not been analyzed in an in vitro cell culture system. In this study, we used the HBV recombinant baculovirus/HepG2 system to measure the time course of antiviral agent-mediated loss of HBV replication as well as the time course and magnitude of HBV production after release from antiviral treatment. Because of the sensitivity of the system, it was possible to measure secreted virions, intracellular replicative intermediates, and nuclear non-protein-bound HBV DNA and separately analyze individual species of DNA, such as single-stranded HBV DNA compared to the double-stranded form and relaxed circular compared to covalently closed circular HBV DNA. We first determined that HBV replication in the HBV recombinant baculovirus/HepG2 system could proceed for at least 35 days, with a 30-day plateau level of replication, making it possible to study antiviral agent-mediated loss of HBV followed by rebound after cessation of drug treatment. All HBV DNA species decreased in a time-dependent fashion following antiviral treatment, but the magnitude of decline differed for each HBV DNA species, with the covalently closed circular form of HBV DNA being the most resistant to drug therapy. When drug treatment ceased, HBV DNA species reappeared with a pattern that recapitulated the initiation of replication, but with a different time course.

scholarly journals Epidemiological and Clinical Features of Hepatitis B Virus Genotypes among Immigrants in Southern Italy

2010 ◽  
Vol 2010 ◽  
pp. 1-6 ◽  
Author(s):  
Gaetano Scotto ◽  
Domenico Martinelli ◽  
Rocco Di Tullio ◽  
Vincenzina Fazio
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Clinical Features ◽  
Hbv Dna ◽  
Hbv Infection ◽  
Hbv Genotypes ◽  
B Virus ◽  
Virus Genotypes ◽  
Severe Chronic Hepatitis

Background/aims. This study aims to determine the distribution and clinical features of HBV-genotypes in a population of immigrants affected by HBV-infection. Methods. Between 01/2003 and 03/2009, 1623 immigrants were tested for HBV-infection. Biochemical and virological activities were determined in HBsAg-positive patients; HBV-genotypes were determined, by the INNO-LiPA HBV Genotyping, in the subjects with HBV DNA detectable. In every patient we evaluated the stage and classified the infection as inactive carrier, mild or moderate/severe chronic hepatitis, cirrhosis, and/or HCC. Results. Among the tested subjects, 191 (11.7%) resulted HBsAg-positive, and in 144/191 (75.4%) serum HBV-DNA was detectable. The genotype distribution was as follows: 45,13% genotype E, 18,1% genotype D, 15,3% genotype B, 13,2% genotype C, 4,9% genotype A, 3,5% mixed genotypes (A–D). The evaluation of liver disease degree showed that 24.6% patients were inactive carriers of HBV infection, 19.4% presented a immunotolerance phase, 34.5% had mild chronic hepatitis, 13.6% had a moderate/severe chronic hepatitis, 6.3% had cirrhosis, and 1.6% presented HCC. Conclusions. Our study evidences a high prevalence of HBV-infection in immigrants, and the potentiality of migratory flow in the introduction of genotype non-D hepatitis B virus. The Hepatitis B virus genotypes presented significant differences in epidemiological and clinical characteristics.

scholarly journals Prevalence of hepatitis B virus genotypes among patients with liver disease in Eritrea

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mohammed Elfatih Hamida ◽  
Saud Mohammed Raja ◽  
Yodahi Petros ◽  
Munir Wahab ◽  
Yemane Seyoum ◽  
...  
Keyword(s):  
Viral Load ◽  
Liver Disease ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hepatitis B Surface Antigen ◽  
Hbv Dna ◽  
Hbv Genotype ◽  
Hbv Genotypes ◽  
B Virus ◽  
Intermediate Endemicity

AbstractEritrea is an East African multiethnic country with an intermediate endemicity for hepatitis B. Our aim was to establish the most prevalent genotypes of hepatitis B virus (HBV) among patients with liver disease. A total of 293 Eritrean patients with liver disease who were hepatitis B surface antigen (HBsAg) positive were enrolled. All sera were tested for liver transaminases, HBV DNA viral load, and hepatitis B seromarkers including HBsAg, anti-HBcAb (total), HBeAg, and anti-HBeAb. Those reactive for HBsAg and anti-HBc (total) were further tested for HBV genotyping. The median (interquartile range) of HBV DNA viral load and ALT levels were 3.47 (1.66) log IU/mL and 28 (15.3) IU/L, respectively. Using type-specific primer-based genotyping method, 122/293 (41.6%) could be genotyped. Irrespective of mode of occurrence, HBV genotype D (21.3%) was the predominant circulating genotype, followed by genotypes C (17.2%), E (15.6%), C/D (13.1%), and C/E (10.7%). Genotypes C/D/E (7.4%), A/D (4.9%), D/E (4.1%), A (2.5%), and B, A/E, B/E, and A/D/C (0.8%) were also present. HBV in Eritrea is comprised of a mixture of HBV genotypes. This is the first study of HBV genotyping among patients with liver disease in Eritrea.

scholarly journals A CRISPR-Cas12b–Based Platform for Ultrasensitive, Rapid, and Highly Specific Detection of Hepatitis B Virus Genotypes B and C in Clinical Application

2021 ◽  
Vol 9 ◽  
Author(s):  
Xu Chen ◽  
Yan Tan ◽  
Shuoshi Wang ◽  
Xueli Wu ◽  
Rui Liu ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Clinical Application ◽  
Point Of Care ◽  
Antiviral Treatment ◽  
Resource Scarcity ◽  
Clinical Samples ◽  
Hbv Genotypes ◽  
B Virus ◽  
Virus Genotypes

Hepatitis B virus (HBV) is one of the most dangerous and prevalent agents that causes acute and chronic liver diseases in humans. Genotyping plays an important role in determining clinical outcomes and response to antiviral treatment in HBV–infected patients. Here, we first devised a CRISPR–based testing platform, termed “CRISPR-HBV,” for ultrasensitive, highly specific, and rapid detection of two major HBV genotypes (HBV-B and HBV-C) in clinical application. The CRISPR-HBV employed multiple cross displacement amplification (MCDA) for rapid preamplification and then Cas12b–based detection for decoding the targets. Finally, the detection result was read out with real-time fluorescence and a lateral flow biosensor. The sensitivity of CRISPR-HBV was 10 copies per test. The specificity was one hundred percent, and no cross reactions were observed in other HBV genotypes and pathogens. The whole detection process, including DNA template extraction (15 min), preamplification reaction of MCDA (30 min at 65°C), CRISPR-Cas12b–based detection (5 min at 37°C), and results readout (∼2 min), could be completed within 1 h. The feasibility of the CRISPR-HBV assay for genotyping HBV-B and -C as successfully validated with clinical samples. Hence, the CRISPR-HBV assay has remarkable potential to develop a point-of-care testing for identifying and distinguishing HBV genotypes B and C in clinical settings, especially in resource-scarcity countries.

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