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.

Colorimetric detection of hepatitis B virus (HBV) DNA based on DNA-templated copper nanoclusters

2016 ◽  
Vol 909 ◽  
pp. 101-108 ◽  
Author(s):  
Xiaoxia Mao ◽  
Siyu Liu ◽  
Chao Yang ◽  
Fengzhen Liu ◽  
Keming Wang ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Colorimetric Detection ◽  
Hbv Dna ◽  
Copper Nanoclusters ◽  
B Virus

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 Characterization of Occult Hepatitis B Virus Infection Among Iranian Patients with Behcet's Disease; Correlation with Clinical Status

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Farhad Shahram ◽  
Saied Ghorbani ◽  
Mahdi Mahmoudi ◽  
Maassoumeh Akhlaghi ◽  
Zohreh Jadali ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Clinical Data ◽  
Disease Duration ◽  
Clinical Manifestations ◽  
Hbv Dna ◽  
Occult Hepatitis ◽  
B Virus ◽  
Occult Hepatitis B ◽  
The Mean

Background: Behcet's disease (BD) is a chronic multisystem vasculitis with an unknown etiology. During the past years, several reports are published on the occult hepatitis B infection (OBI), the presence of hepatitis B virus (HBV) DNA in the absence of HBsAg, in rheumatic diseases. Objectives: The current study aimed to, firstly, investigate the prevalence of OBI in patients with BD, and, secondly, its potential association with the clinical and therapeutic status of BD. Methods: HBV serological markers and HBV DNA were evaluated in 220 consecutive BD patients to detect OBI. Demographic and clinical data of OBI positive and negative groups were compared. Results: The mean age of patients was 39.24 (± 10.57), and 134 (62.9%) were male. The mean disease duration was 14.13 (± 8.63) years. No HBsAg positive case was found, but HBV DNA was found in 19 (8.6%) patients. The median viral load value was 475.84 copy/mL. We compared clinical data of 10 OBI positive and 156 OBI negative BD patients with complete and accessible data. There was no difference between the two groups concerning demographic characteristics (age, sex, and disease duration), different clinical manifestations, or types of medications (immunomodulatory, cytotoxic, and corticosteroids). Conclusions: This is the first study showing a rather high prevalence of OBI among BD patients. We did not find any correlation between OBI positivity and different clinical manifestations, medications, or HLA-B51. Further studies are needed on a larger group of patients and by molecular HBV evaluation (as well as serologic) regarding this possible association.

scholarly journals Comparison of Anti-Hepatitis B Virus Activities of Lamivudine and Clevudine by a Quantitative Assay

2003 ◽  
Vol 47 (1) ◽  
pp. 324-336 ◽  
Author(s):  
Ayman M. Abdelhamed ◽  
Colleen M. Kelley ◽  
Thomas G. Miller ◽  
Phillip A. Furman ◽  
Edward E. Cable ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Antiviral Treatment ◽  
Hbv Dna ◽  
Hbv Infection ◽  
Extracellular Dna ◽  
Quantitative Assay ◽  
Hbv Replication ◽  
B Virus

ABSTRACT In this study, we used a quantitative assay to measure the concentration-dependent effects of antivirals on extracellular hepatitis B virus (HBV) DNA as well as on different cytoplasmic and nuclear forms of HBV DNA that participate in HBV replication. HBV recombinant baculovirus, which efficiently delivers the HBV genome to HepG2 cells, was used for this study because (i) antivirals can be administered prior to initiation of HBV infection or after HBV infection and (ii) sufficiently high HBV replication levels are achieved that HBV covalently closed circular (CCC) DNA can be easily detected and individual HBV DNA species can be quantitatively analyzed separately from total HBV DNA. The results showed that the levels of HBV replicative intermediate and extracellular DNA decreased in a concentration-dependent fashion following antiviral treatment. The 50% effective concentration (EC50) and EC90 values and the Hill slopes differed for the different HBV DNA species analyzed. The data clearly indicated that (i) nuclear HBV DNAs are more resistant to antiviral therapy than cytoplasmic or extracellular HBV DNAs and (ii) nuclear HBV CCC DNA is more resistant than the nuclear relaxed circular form. This report presents the first in vitro comparison of the effects of two antivirals administered prior to initiation of HBV infection and the first thorough in vitro quantitative study of concentration-dependent antiviral effects on HBV CCC DNA.

scholarly journals Semiautomated Quantification of Hepatitis B Virus DNA in a Routine Diagnostic Laboratory

2000 ◽  
Vol 7 (5) ◽  
pp. 853-855 ◽  
Author(s):  
Harald H. Kessler ◽  
Evelyn Stelzl ◽  
Elisabeth Daghofer ◽  
Brigitte I. Santner ◽  
Egon Marth ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Human Error ◽  
Hbv Dna ◽  
Linear Range ◽  
Serum Samples ◽  
Diagnostic Laboratory ◽  
Control Series ◽  
Upper Limit ◽  
B Virus

ABSTRACT The Cobas Amplicor HBV Monitor test for quantitative determination of hepatitis B virus (HBV) DNA in serum has recently been introduced. To evaluate the performance of this assay in a routine diagnostic laboratory, reproducibility of results was determined with the First European Union Concerted Action HBV Proficiency Panel and the Accurun 325 HBV DNA Positive Control, Series 300. Results for 270 routine serum samples were additionally evaluated. To avoid the retesting of a large number of samples due to titers exceeding the upper limit for the linear range of the assay, sera of patients with chronic hepatitis B (CHB) were diluted prior to the assay to 10−4 in normal human plasma, which is included in the assay. The mean coefficient of variation was 22.9% for all input HBV DNAs. Of 270 routine serum samples, 182 (150 sera from transplant donors and 32 sera from patients who had recovered from CHB) tested negative. Eighty-six sera were found to be HBV DNA positive; in six sera, HBV DNA levels were found to exceed the upper limit for the linear range of the assay and had to be retested. In the remaining two sera, inhibition occurred. The semiautomated Cobas Amplicor HBV Monitor test showed sufficient reproducibility and helped in avoiding human error. The relatively narrow linear range of detection is a limitation of the new assay.

scholarly journals Detection of Hepatitis B Virus DNA in Chronic HBV Carriers and Correlation with HBeAg

2017 ◽  
Vol 15 (2) ◽  
pp. 33-36 ◽  
Author(s):  
Shamima Akther ◽  
Md Anwar Husain ◽  
HS Mubarak Hossain
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Real Time ◽  
Real Time Pcr ◽  
Chronic Infection ◽  
Hbv Dna ◽  
B Virus ◽  
Study Population ◽  
Chronic Hbv ◽  
The Mean

Background: Chronic infection with hepatitis B virus causes a spectrum of diseases ranging from asymptomatic infective state to cirrhosis and hepatocellular carcinoma. The detection and quantification of HBV DNA plays an important role in diagnosing HBV infection as well as monitoring therapeutic responses.Methods: The aim of this study was to detect and quantited HBV DNA by real time PCR and to find out correlation HBeAg with HBV DNA in chronic carriers: The present study carried out among 61 previously diagnosed chronic hepatitis B patients.Results: The mean age of study population was 29.20 years. Among the 61 cases, 13(21.3%) were HBeAg positive and 48(78.7%) were HBeAg negative. By real time PCR, DNA detected in 45(73.8%) patients and 16(26.2%) patients were undetected. Association of HBeAg and HBV DNA was observed in 13 HBeAg positive cases, where 12(92.3%) had detectable DNA and in 48 HBeAg negative patients, 33(68.8%) had detectable DNA while 15(31.2%) were undetected. The present study observed a higher viral load (105copies/ml) among HBeAg positive patients (84.6%) than HBeAg negative patients (12.5%).Conclusion: The present study observed that, there was positive correlation among HBeAg and HBV DNA in chronic HBV carriers. However, there was some discordance observed among HBeAg and HBV DNA. Therefore, in addition to HBeAg, HBV DNA should be assessed for appropriate evaluation of CHB carriers.Chatt Maa Shi Hosp Med Coll J; Vol.15 (2); Jul 2016; Page 33-36

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.

272 Use of LioCyx-M, autologous hepatitis B virus (HBV)-Specific T cell receptor (TCR) T-cells, in advanced HBV-related hepatocellular carcinoma (HCC)

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A297-A297
Author(s):  
Fu-Sheng Wang ◽  
Fanping Meng ◽  
Jiehua Jin ◽  
Yuanyuan Li ◽  
Regina Wanju Wong ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
T Cells ◽  
Hepatitis B Virus ◽  
T Cell ◽  
Hepatitis B ◽  
T Cell Receptor ◽  
Dose Level ◽  
Cell Receptor ◽  
Hbv Dna ◽  
B Virus

BackgroundWe have demonstrated the ability of Hepatitis-B-virus (HBV)-specific T cell receptor (TCR) bioengineered T cells to recognize and lyse Hepatocellular carcinoma (HCC) cells expressing HBV antigens derived from HBV-DNA integration in patients with liver transplant.1 LioCyx-M is an immunotherapeutic product composing of autologous T cells transiently modified with in-vitro transcribed mRNA encoding HBV-specific TCR. The transient TCR expression makes LioCyx -M amenable to a dose escalating posology.MethodsThe primary endpoint of this phase 1 trial is to assess the safety and tolerability of LioCyx-M in patients with advanced HBV-HCC without curative treatment options. Eligible patients were diagnosed with Barcelona clinic liver cancer stage B or C HCC (Child-Pugh < 7 points), receiving >1 year antiviral treatment prior to enrollment. These patients had matching HLA class I genotypes which present HBV encoded antigen. Peripheral blood was collected from each patient prior to each dose for LioCyx-M manufacturing. Patients received 4 escalating doses of 1×104 cells/kg, 1×105 cells/kg, 1×106 cells/kg, 5×106 cells/kg bodyweight (BW) in the first treatment cycle, each intravenously administered weekly. Patients underwent 1-month safety assessment post the 4th infusion, according to Common Terminology NCI CTCAE Version 4.0.3. If there were no dose associated toxicities, patients were eligible to continue administration of LioCyx-M at dose of 5 × 106 cells/kg BW weekly. Tumor response per RECIST 1.1 criteria and survival time were assessed.ResultsAt data cutoff (30 April 2020), eight patients were enrolled, with a median age of 53 (range: 49 - 67). These patients received a median number of 6 (range: 4 - 12) infusions of LioCyx-M. 1 patient developed Grade 3 elevations in alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST) and bilirubin after receiving LioCyx-M at dose level of 1×105 cells/kg BW. Another patient had Grade 1 transient fever after receiving LioCyx-M at dose level 5×106 cells/kg BW in the 4th, 5th and 6th infusions. No treatment-related adverse events (trAEs) such as cytokine release syndrome or neurotoxicity were observed. No fatal trAEs were observed. The median time to progression was 1.9 months (range: 0.2 - 9.5 months). The median overall survival was 34 months (range: 3 - 38.2 months).ConclusionsThe encouraging clinical outcome and tolerable safety highlight the good benefit-risk profile of LioCyx-M. Therefore, further exploration of efficacy of LioCyx-M treatment for advanced HBV-HCC is warranted in a Phase 2 proof-of-concept clinical study.AcknowledgementsFunding: Lion TCR.Trial RegistrationNCT03899415Ethics ApprovalThe study was approved by Fifth Medical Center of Chinese PLA General Hospital’s Ethics Board, approval number R2016185DI010.ReferenceTan AT, Yang N, Lee Krishnamoorthy T, et al. Use of Expression Profiles of HBV-DNA Integrated Into Genomes of Hepatocellular Carcinoma Cells to Select T Cells for Immunotherapy. Gastroenterology 2019;156(6):1862–1876.e9.

scholarly journals A Comparison Research of Hepatitis B Virus Large Surface Protein with HBV DNA Detecting

2008 ◽  
Vol 12 ◽  
pp. e415
Author(s):  
Z.L. Wu ◽  
X.D. Lu ◽  
X.Q. Zhong ◽  
L.F. Ling ◽  
G. Lin ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Protein ◽  
Hbv Dna ◽  
Comparison Research ◽  
B Virus

scholarly journals Occult Hepatitis B Virus Infection among HIV Positive Patients in Nigeria

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Oluyinka Oladele Opaleye ◽  
Adeolu Sunday Oluremi ◽  
Adetona Babatunde Atiba ◽  
Moses Olubusuyi Adewumi ◽  
Olatunji Victor Mabayoje ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Virus Infection ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Hiv Positive ◽  
Hepatitis B Virus Infection ◽  
Occult Hepatitis ◽  
B Virus ◽  
Occult Hepatitis B ◽  
Detectable Hbsag

HIV has been known to interfere with the natural history of hepatitis B virus (HBV) infection. In this study we investigate the prevalence of occult hepatitis B virus infection (OBI) among HIV-infected individuals in Nigeria. Overall, 1200 archived HIV positive samples were screened for detectable HBsAg using rapid technique, in Ikole Ekiti Specialist Hospital. The HBsAg negative samples were tested for HBsAg, anti-HBc, and anti-HCV by ELISA. Polymerase chain reaction was used for HBV DNA amplification and CD4 counts were analyzed by cytometry. Nine hundred and eighty of the HIV samples were HBsAg negative. HBV DNA was detected in 21/188 (11.2%) of patients without detectable HBsAg. CD4 count for the patients ranged from 2 to 2,140 cells/μL of blood (mean = 490 cells/μL of blood). HCV coinfection was detected only in 3/188 (1.6%) of the HIV-infected patients (P>0.05). Twenty-eight (29.2%) of the 96 HIV samples screened were positive for anti-HBc. Averagely the HBV viral load was <50 copies/mL in the OBI samples examined by quantitative PCR. The prevalence of OBI was significantly high among HIV-infected patients. These findings highlight the significance of nucleic acid testing in HBV diagnosis in HIV patients.

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