Abundance of Noncircular Intrahepatic Hepatitis B Virus DNA May Reflect Frequent Integration Into Human DNA in Chronically Infected Patients

Abstract Background Hepatitis B virus (HBV) integration has implications for cancer development and surface antigen (HBsAg) production, but methods to quantify integrations are lacking. The aim of this study was to develop a droplet digital PCR (ddPCR) assay discriminating between circular and integrated HBV DNA, and to relate the distribution between the two forms to other HBV markers. Methods ddPCR with primers spanning the typical linearization breakpoint in the HBV genome allowed for quantification of the absolute copy numbers of total and circular HBV DNA, and calculation of linear HBV DNA. Results Analysis of 70 liver biopsies from patients with chronic HBV infection revealed that the fraction of linear HBV DNA, which includes integrations, was higher in HBeAg-negative patients than HBeAg-positive. The ratio between HBsAg and HBV DNA levels in serum correlated with the intrahepatic proportion of linear HBV DNA. Furthermore, ddPCR experiments on serum samples and experiments with nuclease indicated the contribution of encapsidated double-stranded linear DNA and replication intermediates to be limited. Conclusions The degree of integration of intrahepatic HBV DNA in the HBeAg-negative stage may be higher than previously anticipated, and integrated DNA may explain the persistence of high HBsAg serum levels in patients with low HBV DNA levels.

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Diagnostic Value of Detection of Pregenomic RNA in Sera of Hepatitis B Virus-Infected Patients with Different Clinical Outcomes

Journal of Clinical Microbiology ◽

10.1128/jcm.01275-19 ◽

2019 ◽

Vol 58 (2) ◽

Cited By ~ 1

Author(s):

Ni Lin ◽

Aizhu Ye ◽

Jinpiao Lin ◽

Can Liu ◽

Jinlan Huang ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Clinical Outcomes ◽

Hepatitis B Surface Antigen ◽

Surrogate Marker ◽

Diagnostic Value ◽

Hbv Dna ◽

Hbv Cccdna ◽

B Virus ◽

Intrahepatic Hbv

ABSTRACT Pregenomic RNA (pgRNA) is a direct transcription product of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), and it plays important roles in viral genome amplification and replication. This study was designed to investigate whether serum pgRNA is a strong alternative marker for reflecting HBV cccDNA levels and to analyze the correlation between serum pgRNA, serum HBV DNA, and hepatitis B surface antigen (HBsAg). A total of 400 HBV-infected patients who received nucleos(t)ide analog (NA) therapy with different clinical outcomes were involved in this research. Case groups included asymptomatic hepatitis B virus carrier (ASC), chronic hepatitis B (CHB), liver cirrhosis (LC), and hepatocellular carcinoma (HCC) patients, with 100 patients in each group. The results showed that the levels of HBV pgRNA had significant differences between these 4 groups. Serum pgRNA levels correlated well with serum HBV DNA and HBsAg levels (HBV pgRNA levels versus HBV DNA levels, r = 0.58, P < 0.001; HBV pgRNA levels versus HBsAg levels, r = 0.47, P < 0.001). In addition, we focused on the 108 HBV-infected patients with HBV DNA levels of <500 IU/ml; it was surprising to find that in 17.57% (13/74) of cases, HBV pgRNA could be detected even when the HBV DNA level was below 20 IU/ml. In conclusion, HBV pgRNA levels in serum can be a surrogate marker for intrahepatic HBV cccDNA compared with serum HBV DNA and HBsAg. The detection of serum HBV pgRNA levels may provide a reference for clinical monitoring of cccDNA levels and the selection of appropriate timing for discontinuing antiviral therapy, especially when HBV DNA levels are below the detection limit.

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Pradefovir* appears to yield clinically significant reductions in serum levels of Hepatitis B virus (HBV) DNA,

Inpharma Weekly ◽

10.2165/00128413-200514860-00014 ◽

2005 ◽

Vol &NA; (1486) ◽

pp. 7

Author(s):

&NA;

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Serum Levels ◽

Hbv Dna ◽

B Virus ◽

Clinically Significant

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PREVENTION OF HEPATITIS B VIRUS CARRIER STATE IN INFANTS ACCORDING TO MATERNAL SERUM LEVELS OF HBV DNA

The Lancet ◽

10.1016/s0140-6736(89)90003-2 ◽

1989 ◽

Vol 333 (8635) ◽

pp. 406-410 ◽

Cited By ~ 88

Author(s):

HenriettaM.H. Ip ◽

VivianC.W. Wong ◽

P.Nico Lelie ◽

MaryC. Kuhns ◽

HenkW. Reesink

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Serum Levels ◽

Carrier State ◽

Hbv Dna ◽

Maternal Serum ◽

B Virus ◽

Hepatitis B Virus Carrier ◽

Virus Carrier

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Correlation of HBcrAg with Intrahepatic Hepatitis B Virus Total DNA and Covalently Closed Circular DNA in HBeAg-Positive Chronic Hepatitis B Patients

Journal of Clinical Microbiology ◽

10.1128/jcm.01303-18 ◽

2018 ◽

Vol 57 (1) ◽

Cited By ~ 5

Author(s):

Lin Wang ◽

Xi Cao ◽

Zhenzi Wang ◽

Yuhua Gao ◽

Juan Deng ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Hbv Dna ◽

Circular Dna ◽

Hbv Cccdna ◽

B Virus ◽

Positive Chronic Hepatitis ◽

Total Dna ◽

Intrahepatic Hbv

ABSTRACT The purpose of this study was to explore the correlations of serum hepatitis B core-related antigen (HBcrAg) with intrahepatic Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) and HBV total DNA in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. Serum HBcrAg and other parameters, including HBV DNA, HBV RNA, HBeAg, hepatitis B surface antigen (HBsAg), hepatitis B core antibody (anti-HBc), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively measured at baseline and follow-up time points. Intrahepatic HBV cccDNA and total DNA were quantitatively detected at baseline and 96 weeks. Grading of liver necroinflammation and staging of hepatic fibrosis were assessed at baseline and 96 weeks. Correlations between serum HBcrAg and other parameters were analyzed by Pearson’s correlation analysis. The results showed that pretreatment HBcrAg correlated significantly with HBV total DNA levels (r = 0.328, P = 0.003) in 82 CHB patients, and, after removing three outliers, with intrahepatic HBV cccDNA (r = 0.323, P = 0.004; n = 79). Serum HBcrAg correlated better with HBV cccDNA in patients with lower levels of serum HBV DNA (stratified by 7 log IU/ml of HBV DNA; r = 0.656, P = 0.003 versus r = −0.02, P = 0.866). Significant inverse correlations were found between HBcrAg and grade of liver necroinflammation (r = −0.245, P = 0.037), stage of hepatic fibrosis (r = −0.360, P = 0.002) at baseline. Serum HBcrAg presented significant correlation with intrahepatic HBV cccDNA in patients with HBeAg seroconversion at 96 weeks (r = 0.622, P = 0.006). The decrease in HBcrAg showed significant correlation with the decrease in HBV cccDNA after 96-week NA therapy (r = 0.282, P = 0.043). Serum HBcrAg also correlated significantly with other serum markers at baseline and 96 weeks of NA therapy. In conclusion, baseline HBcrAg and its decreased value were significantly correlated with the corresponding intrahepatic HBV cccDNA.

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Mechanisms downstream of reverse transcription reduce serum levels of HBV DNA but not of HBsAg in chronic hepatitis B virus infection

Virology Journal ◽

10.1186/s12985-015-0447-5 ◽

2015 ◽

Vol 12 (1) ◽

Cited By ~ 11

Author(s):

Simon B. Larsson ◽

Sebastian Malmström ◽

Charles Hannoun ◽

Gunnar Norkrans ◽

Magnus Lindh

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B Virus ◽

Virus Infection ◽

Hepatitis B ◽

Reverse Transcription ◽

Serum Levels ◽

Hbv Dna ◽

Hepatitis B Virus Infection ◽

Chronic Hepatitis B Virus ◽

B Virus

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Evaluation of Serum Levels of HBV-DNA Concentration and HBSAG Titers of Hepatitis B Virus-Infected Subjects at NAUTH Nnewi

Saudi Journal of Medicine ◽

10.36348/sjm.2020.v05i01.006 ◽

2020 ◽

Vol 05 (01) ◽

pp. 25-31

Author(s):

Obiomah Chinwe Favour ◽

Amilo Grace I ◽

Kalu Stephen O ◽

Ndulue Israel N ◽

Obeagu Emmanuel Ifeanyi

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Serum Levels ◽

Hbv Dna ◽

Dna Concentration ◽

B Virus

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Distinct Hepatitis B Virus Dynamics in the Immunotolerant and Early Immunoclearance Phases

Journal of Virology ◽

10.1128/jvi.02164-09 ◽

2010 ◽

Vol 84 (7) ◽

pp. 3454-3463 ◽

Cited By ~ 64

Author(s):

Hurng-Yi Wang ◽

Ming-Hung Chien ◽

Hsiang-Po Huang ◽

Hsiao-Chi Chang ◽

Chung-Che Wu ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Early Stage ◽

Host Immunity ◽

Viral Population ◽

Dominant Factor ◽

Serum Samples ◽

Copy Numbers ◽

Viral Copy Number ◽

B Virus

ABSTRACT Little is known about hepatitis B virus (HBV) diversity changes within a host during the immunotolerant phase of chronic HBV infection. Such knowledge, nevertheless, may help in understanding how host immunity and HBV interact at the early stage of infection. In this study, serial serum samples were collected from a long-term (>17 years) follow-up cohort of seven patients, and multiple copies of the full-length viral genome from serially sampled sera were recovered and analyzed. Viral genetic diversity was positively correlated with host immunity, represented by levels of alanine aminotransferase (ALT), but was negatively correlated with the viral copy number. During the immunotolerant phase, when the host immunity was feeble (ALT < 20 U/liter), viral nucleotide diversity decreased while copy numbers increased. Rates of evolutionary change derived for different patients were in a very narrow range (1.6 × 10−5 to 5.4 × 10−5/site/year). As the disease progressed toward the immunoclearance phase (ALT > 20 U/liter), viral diversity increased but copy numbers decreased. Evolutionary rates varied among patients in accordance with their levels of ALT, ranging from 9.6 × 10−6 to 3.2 × 10−4/site/year. More than half (19/32 sites) of positively selected sites resided in immune epitopes, suggesting their possible role in host immunity. Our results demonstrate that host immunity is a dominant factor in HBV evolution. Different selective forces, including immune-mediated positive selection and virus-mediated negative selection, operate in tandem in shaping viral population dynamics within a host.

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

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.7.5.853-855.2000 ◽

2000 ◽

Vol 7 (5) ◽

pp. 853-855 ◽

Cited By ~ 7

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.

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