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Characterization of the hepatitis B virus DNA detected in urine of chronic hepatitis B patients.

ABSTRACT: BACKGROUND:Detection of human hepatitis B virus (HBV) DNA in the urine of patients with chronic hepatitis B infection (CHB) has been reported previously, suggesting urine could provide a potential route of horizontal HBV transmission. However, it is not clear whether the HBV DNA detected in urine is indeed full-length, infectious viral DNA. The aim of this study is to assess the potential infectivity of urine from patients with CHB and to correlate HBV DNA detection in urine with clinical parameters, such as serum viral load and HBeAg status. METHODS:Urine from 60 CHB patients with serum viral loads ranging from undetectable to 108 IU/mL were analyzed for HBV DNA and serum immune markers. HBV DNA was detected from total urine DNA and size-fractionated urine DNA (separated into ≤1 kb and > 1 kb fractions) by PCR analysis of six regions of the HBV genome. RESULTS:Twenty-seven of 59 (45.7%) patients with HBV serum viral load (≥20 IU/mL) contained at least 20 copies per mL of fragmented HBV DNA in urine detected in at least 1 of the 6 PCR assay regions. Only one patient contained HBV DNA detected by all six regions, and was found to have evidence of blood in the urine. Sixteen of 25 urine samples with high viral load (> 105 IU/mL) and 11 of 34 urine samples with low viral load (< 105 IU/mL) contained detectable HBV DNA. Twelve of 27 (44.44%) patients with detectable HBV DNA in urine were HBeAg positive, and only 5 of these HBeAg positive patients were in the group of 33 (15.15%) patients with no detectable HBV DNA in urine. By Fishers' exact test, HBV DNA in urine is significantly associated with high serum viral load (P = 0.0197) and HBeAg (P = 0.0203). CONCLUSIONS:We conclude that urine from CHB patients with healthy kidney function should not contain full-length HBV DNA, and therefore should not be infectious.

SUBMITTER: Jain S

PROVIDER: S-EPMC5857095 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundChronic Hepatitis B infection is a major health problem in Republic of Congo therefore molecular analysis of HBV strains is important to detect the patients at high risk of disease progression.MethodsSerum samples were obtained from 111 chronic HBV patients in Pointe Noire. HBsAg, HBeAg and HBeAb were detected. A fragment of the preS1 region of HBV was amplified and sequenced to determine genotypes, subgenotypes and to identify mutations.ResultsOf the 111 samples analyzed, 35 patients were asymptomatic carriers (ASC), 24 with a chronic active hepatitis (CAH), 33 with liver cirrhosis (LC) and 19 have a hepatocellular carcinoma (HCC). The mean age were 45 ± 13 year, 88 (79.3 %) were male and 23 (20.7 %) female. The prevalence of HBeAg was 15.3 % and 73 % of subjects were anti-HBe positive. The mean serum level of alanine aminotransferase transaminase (ALT) and aspartate transaminase (AST) was 25.1 ± 9 IU/L and 28.6 ± 10 IU/L respectively. Eighty two samples out of 111 (73.9 %) were genotyped by the analyzing of the S region of HBV, 58 (70.7 %) cases belonged to HBV genotype E and 24 (29.3 %) were genotype A with three subgenotypes; A3 (66.7 %), A4 (20.8 %) and A6 (12.5 %). Prevalence of genotype A was relatively high in CAH (33.3 %) and HCC (31.6 %) patients in comparison with other groups. The most prevalent amino acids substitutions were R38K found in 14 (17.1 %) sequences, following by H44L in 11 (13.4 %), K13E in 8 (9.8 %), N29K in 8 (9.8 %), A35E in 8 (9.8 %), V80I in 7 (8.5 %) and in 6 (7.3 %) sequences for S90T. Different substitutions located in the hepatocyte binding site were higher among patients with LC and HCC (p < 0.05).ConclusionsThis study have shown that HBV genotype E and A were the most frequent strains circulating in Republic of Congo patients. HBV pres1 substitutions found in this study were associated with severe clinical forms of liver diseases. This data have shown the importance of implementing an effective program to fight HBV infection.

Project description: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.

Project description:Purpose: Worldwide, over 350 million people are chronically infected with the hepatitis B virus (HBV) and are at increased risk of developing progressive hepatic diseases (hepatitis, cirrhosis or liver cancer). Still, the metabolic processes operational during the distinct clinical phases of a chronic HBV infection to liver disease remains unexplored. Thus, the aim of our study was to determine the serum metabolic characteristics of hepatic disease patients with chronic hepatitis B in active and inactive stage and study the influence of active and inactive stage of the chronic hepatitis B virus on the development of liver cancer by untargeted metabolomics. Method: To investigate this, we conducted an untargeted metabolomics approach to determine the metabolic characteristics of 199 human serum samples from hepatic disease patients in active and inactive chronic HBV via UHPLC-Q-TOF/MS, and patients with HBV in active or inactive stage were grouped based on their serum levels of HBV DNA and HBV envelope antigen (HBeAg). Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and volcano plot were used for multivariate analysis. Pathway analysis were conducted according to p-values from pathway enrichment analysis and pathway impact values from pathway topology analysis. In addition, commercial databases were used to identify metabolites from significantly altered metabolic pathways. All samples were analyzed for liver function tests indicated the levels of certain enzymes. Results: Our data provide insight into the metabolic dysregulation experienced during active or inactive stage of the HBV on the development of liver disease, and analysis of serum metabolomic profiles indicated that most of the altered metabolic pathways for chronic hepatitis and cirrhosis patients with active or inactive stage of the HBV were associated with D-Glutamine and D-glutamate metabolism, indicating a strong link to HBV replication (p < 0.05). In addition, liver function analysis showed the enzymes gamma-glutamyl transferase (GGT) were increased concomitant to the progression of hepatitis to cirrhosis induced by chronic HBV (p < 0.05).

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Characterization of the hepatitis B virus DNA detected in urine of chronic hepatitis B patients.

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