BACKGROUND AND AIMS:Chronic hepatitis D is the most severe form of viral hepatitis. Despite recommendations for systematic screening of HBsAg-positive individuals, hepatitis D infection remains underdiagnosed. Standard HDV virological markers are classically assessed in serum or plasma specimens obtained through venous whole blood sampling. This approach is costly and challenging in resource-limited settings. Dried blood spot (DBS) sampling offers a practical alternative for large-scale screening, diagnosis, and monitoring of viral hepatitis. The goal of the study is to evaluate the performance of commercially available HDV diagnostic assays using DBS. METHODS:Plasma and DBS-collected whole blood specimens from individuals chronically infected with HBV or coinfected with HBV and HDV were analysed for HDV antibody detection, HDV RNA quantification, HBsAg levels, and HDV genotyping. RESULTS:HDV antibodies were reliably detected in DBS after threshold adjustment, with the LIAISON XL Murex Anti-HDV assay showing superior sensitivity (99.3%) compared to HDV Ab DIA.PRO (90.3%), while both assays exhibited excellent specificity. HDV RNA was quantifiable in 86.8% of DBS specimens from patients with active infection, although levels were about 1.5 log10 lower than plasma, showing a strong correlation (r = 0.786; p < 0.0001). HBsAg was detectable and quantifiable in all DBS samples, correlating closely with plasma values (r = 0.98). DBS-based HDV genotyping was successful in 84.8% of samples and concordant with plasma results. CONCLUSIONS:DBS collection of whole blood from DBS appears to be a promising and practical alternative to conventional venous blood sampling for HDV screening, diagnosis, and monitoring.

Background:Superinfection with the hepatitis D virus (HDV) leads to a more aggressive form of chronic hepatitis B. While around 5% of hepatitis B surface antigen (HBsAg)-positive individuals are estimated to be hepatitis B virus (HBV)-HDV dually infected globally, the time point of superinfection is unknown and repeated HDV testing is not yet supported by international guidelines. Inci-D is a post hoc analysis from 2 prospective cohorts to evaluate the HDV superinfection rate. Method:The Inci-D cohort consists of 2 HBV cohorts of clinical meta-data and stored plasma samples or dried-blood spots (DBS) from The Gambia (Prolifica) and France. Results:Overall, samples from 1016 HBsAg-positive individuals were analyzed (625 from The Gambia, 391 from France); the baseline HDV prevalence was 1.1% (7/625) and 2.5% (10/391), respectively. The median age (interquartile range) was 38 (32-50) years, and 63% were male. Patients in the French cohort were older (P < .001), with higher liver enzymes (P < .001), were more often hepatitis B e antigen positive (P < .001) or anti-HCV positive (P < .001), and had more advanced liver disease (P < .001). In the Gambian cohort, after a median follow-up time of 5.98 years, 14 individuals were detected to be newly HDV antibody (Ab) positive (3.85/1000 patient-years). In the French cohort, after a median follow-up time of 2.1 years, 3 individuals were newly detected to be HDV Ab positive (3.70/1000 patient-years). Conclusions:Hepatitis Delta superinfection increases considerably in HBsAg-positive carriers in The Gambia as well as in France. These findings support consideration of repeated HDV serology testing in HBsAg-positive individuals.

BACKGROUND & AIMS:Chronic hepatitis B (CHB) and metabolic dysfunction-associated steatotic liver disease (MASLD) commonly co-exist, with conflicting data in prevalence and disease severity. We aimed to investigate these discrepancies. METHODS:This multicenter study included consecutive patients with CHB from 19 European centers. A survey on standard of care for MASLD screening in CHB was circulated. RESULTS:A total of 1709 patients with CHB were included; median age, 53 years (interquartile range [IQR], 42-64); males, 60.7%; body mass index (BMI), 25.6 kg/m2 (IQR, 14-63 kg/m2); and 57.3% White. MASLD prevalence (1510 consecutive patients) was 42.3%. BMI (odds ratio [OR], 1.27; 95% confidence interval [CI], 1.19-1.36), ferritin (OR, 1.00; 95% CI, 1.00-1.00) and type 2 diabetes (OR, 2.60; 95% CI, 1.12-6.02) were independently associated with MASLD. The prevalence of advanced fibrosis was 18% (255/1420) in the whole cohort, 25.4% (162/639) among patients with CHB with MASLD, and 13.7% in those without MASLD. Independent predictors of advanced fibrosis were MASLD (OR, 2.76; 95% CI, 1.50-5.05), BMI (OR, 1.08; 95% CI, 1.02-1.15), alanine transaminase (OR, 1.01; 95% CI, 1.00-1.03), lower platelets (OR, 0.99; 95% CI, 0.98-0.99), insulin treatment (OR, 13.88; 95% CI, 2.95-65.28), and long-term antivirals (OR, 4.86; 95% CI, 2.40-9.85). During follow-up (48 months), only patients without MASLD showed significant liver stiffness measurement improvement over time (P < .001). Among patients with MASLD, Fibrosis-4 and liver stiffness measurement performed moderately at predicting advanced fibrosis (area under the receiver operating characteristic curve = 0.71 vs 0.70; P = .38) against histology. As standard of care, 68.4% of centers screened all patients with CHB for MASLD; 52.6% followed the same treatment indication in those with CHB and MASLD vs CHB only. CONCLUSIONS:In this large European cohort, MASLD and fibrosis were highly prevalent among patients with CHB, whereas MASLD aggravated liver fibrosis. Though screening strategies remain inconsistent, ferritin levels, increased BMI, and type 2 diabetes may inform on the presence of MASLD. Biomarkers showed modest performance in predicting fibrosis.

Background Treatment of chronic hepatitis B infection requires lifelong administration of nucleos(t)ide analogues with a high barrier to resistance and effective viral suppression. The major limitation of lifelong therapy is the possible selection of drug-resistant hepatitis B virus (HBV) strains. International Liver Society guidelines recommend that hepatitis B resistance testing must be performed by a reference laboratory.Objectives Performance of the deep sequencing-based ViroKey (R) SQ FLEX Genotyping Assay for the determination of HBV genotypes and resistance profiles were evaluated.Patients and methods Plasma samples collected from patients with chronic hepatitis B have been sequenced by two methods including Sanger (population) sequencing of a portion of the P/S overlapping gene and the deep sequencing-based ViroKey (R) SQ FLEX Genotyping Assay (Vela Diagnostics).Results A high concordance rate with population sequencing was found regardless of HBV genotypes. Deep sequencing with the Sentosa platform was more sensitive than population sequencing in detecting minor variant populations.Conclusions The deep sequencing-based ViroKey (R) SQ FLEX Genotyping Assay can be confidently used in clinical practice for hepatitis B genotyping and resistance testing.

Measurement of hepatitis B virus (HBV) DNA levels is the standard of care for diagnosis active HBV infection, assessing disease severity and prognosis, and guiding treatment decisions and monitoring response to therapy. In the study, the analytical and clinical performance of the ELITe InGenius System for quantifying HBV DNA was evaluated. A total of 377 of archived EDTA plasma or serum specimens were tested. The HBV ELITe MGB Kit demonstrated linear quantification of HBV DNA with high precision. The specificity was 100 %. There was a satisfactory correlation with the cobas HBV Test, including patients on antiviral treatment with nucleos(t)ide analogues. The HBV ELITe MGB Kit using the ELITe InGenius System is suitable to detect and quantify HBV DNA in HBsAg-positive patients across the entire quantification dynamic range.

Hepatitis B and C infections are an underdiagnosed global health problem. Measurement of HBV DNA or HCV RNA levels using nucleic acid-based molecular diagnostic assays has been established as the standard of care for assessing diagnosis, guiding the treatment decision, and evaluating responses to antiviral therapy. In the present study, we examined the performance of the cobas 5800 System for HBV DNA and HCV RNA quantification in a large series of patients chronically infected. Specificity of the cobas HBV and HCV Tests on the 5800 System was high (99.1 % and 100 %, respectively). Linearity using the AcroMetrix panels was excellent. Repeatability and intermediate precision coefficients of variation were within 5 %. Of the 334 clinical specimens tested in parallel on the cobas 5800 and cobas 4800 Systems for HBV and the m2000 RealTime or Alinity m Systems for HCV, only 12 (3.6 %) yielded discrepant results that were at or near the limit of quantification of the cobas 5800 assays. The correlation between viral load results was extremely high, and only weak bias were observed across the entire range of concentrations tested without clinical impact in patients who are eligible for antiviral therapy. This comparison study demonstrated equivalent performance of the new cobas 5800 System compared with other molecular platforms widely used in clinical practice for HBV DNA and HCV RNA quantification. The cobas 5800 System can be confidently used in clinical practice. A few clinical specimens with low viral loads may be missed. Further studies are warranted to confirm or refute this finding.

The ideal non-invasive marker to identify patient at risk for liver fibrosis in the general population is unknown. Current guidelines (EASL) recommend using the FIB-4 score as a screening tool but the low specificity may result in a large number of potentially false-positive results. Recently, the liver risk score (LRS) has been suggested to more accurately identify patients at-risk. The aim of our study was to identify populations at-risk using the two scores. “Cerbafib” was a prospective cohort derived from laboratories throughout the Paris region. The prevalence of advanced liver fibrosis was assessed using the non-invasive FIB-4 score with a cut-off of 2.67 (rule-in) and 1.3 (rule-out) and the LRS with a cut-off of < 6 (minimal risk), ≥ 6 to < 10 (low risk), ≥ 10 to < 15 (medium risk) and ≥ 15 (high risk). Between January and April 2023, 179 865 patients were included in the cohort. The mean age was 52 years and 45

OBJECTIVES:To identify sexual/sex-associated risk factors for hepatitis C transmission among men who have sex with men (MSM) and visualise behavioural trajectories from 2019 to 2021. METHODS:We linked a behavioural survey to a hepatitis C cohort study (NoCo), established in 2019 across six German HIV/hepatitis C virus (HCV) treatment centres, and performed a case-control analysis. Cases were MSM with recent HCV infection, and controls were matched for HIV status (model 1) or proportions of sexual partners with HIV (model 2). We conducted conditional univariable and multivariable regression analyses. RESULTS:In all, 197 cases and 314 controls completed the baseline questionnaire and could be matched with clinical data. For regression models, we restricted cases to those with HCV diagnosed since 2018 (N = 100). Factors independently associated with case status included sex-associated rectal bleeding, shared fisting lubricant, anal douching, chemsex, intravenous and intracavernosal injections, with population-attributable fractions of 88% (model 1) and 85% (model 2). These factors remained stable over time among cases, while sexual partner numbers and group sex decreased during COVID-19 measures. CONCLUSIONS:Sexual/sex-associated practices leading to blood exposure are key factors in HCV transmission in MSM. Public health interventions should emphasize the importance of blood safety in sexual encounters. Micro-elimination efforts were temporarily aided by reduced opportunities for sexual encounters during the COVID-19 pandemic.

BACKGROUND:Expanding antiviral therapy to people with chronic hepatitis B virus (HBV) infection who are ineligible to receive treatment under current international criteria has been increasingly debated. Evidence to support this approach is scarce, especially in Africa. We aimed to address this knowledge gap by analysing the clinical outcomes of people with chronic hepatitis B in The Gambia who were untreated and ineligible for antiviral therapy at diagnosis. METHODS:Between Dec 7, 2011, and Jan 24, 2014, we implemented the prospective PROLIFICA cohort study in The Gambia. Participants with chronic hepatitis B aged 16 years or older were recruited after large-scale, community-based HBV screening; blood bank-based HBV screening in Edward Francis Small Teaching Hospital, Banjul; and prospective follow-up of HBsAg-positive individuals via historical, population-based HBsAg serosurveys in two rural villages (Keneba and Manduar). Participants underwent HBV serology and other laboratory tests, fasting FibroScan, and abdominal ultrasound. Survival data were collected between Dec 7, 2011, and Aug 17, 2021. Between Oct 9, 2018, and Aug 17, 2021, all HBsAg-positive participants enrolled in the 2011-14 cohort were invited for a reassessment. For this analysis, we included HBsAg-positive people and excluded all participants who were eligible for treatment according to the 2012 European Association for the Study of the Liver (EASL) criteria at baseline and those who were treated irrespective of treatment eligibility. The primary outcome was all-cause mortality, assessed in all treatment-ineligible and treatment-naive participants with follow-up data. The secondary outcome, analysed in those who were reassessed, was disease progression, defined as becoming eligible for antivirals per 2017 EASL criteria; having an increase in liver fibrosis of at least one stage; or having a clinical diagnosis of hepatic decompensation or hepatocellular carcinoma. FINDINGS:943 HBsAg-positive people with chronic hepatitis B were recruited to the PROLIFICA study. Of these 943, 58 (6%) fulfilled 2012 EASL treatment eligibility criteria at baseline, 35 (4%) were ineligible for treatment but received antiviral therapy, and 44 (5%) were immediately lost to follow-up. Thus, 806 (85%) participants were analysed for the primary outcome (486 [60%] were male and 320 [40%] were female). After a median follow-up of 6·11 years (IQR 5·34-6·80), 708 (88%) participants were confirmed to be alive at last surveillance, 71 (9%) were lost to follow-up and were censored, and 27 (3%) died, giving an all-cause mortality rate of 582 per 100 000 person-years (95% CI 399-849). Of the 27 people who died, five (19%) had liver-related deaths. Of 708 participants confirmed to be alive, 544 (77%) attended follow-up and were assessed for the secondary outcome. Disease progression occurred in 36 (7%) participants: five (1%) became newly eligible for antiviral therapy per EASL 2017 criteria without liver fibrosis progression; 18 (3%) had liver fibrosis progression alone; 13 (2%) had liver fibrosis progression and newly fulfilled the treatment criteria; and none had hepatic decompensation or developed hepatocellular carcinoma. In multivariable analysis adjusted for sex and age, only a baseline HBV DNA of 20 000 IU/mL or more, compared with the baseline HBV DNA of 2000 IU/mL or lower as the reference, was significantly associated with liver disease progression (odds ratio 5·39, 95% CI 1·37-21·23). INTERPRETATION:Among people with chronic hepatitis B who were ineligible for antiviral therapy in The Gambia, all-cause mortality and liver disease progression were low. The clinical benefit of expanding antiviral therapy in this subgroup of patients remains uncertain. FUNDING:European Commission, Medical Research Council UK Research and Innovation, and Gilead Sciences.

BACKGROUND AND AIMS:Chronic hepatitis delta represents a major global health burden. Clinical features of hepatitis D virus (HDV) infection vary largely between different regions worldwide. Treatment approaches are dependent on the approval status of distinct drugs and financial resources. METHODS:The Hepatitis Delta International Network (HDIN) registry involves researchers from all continents (Wranke, Liver International 2018). We here report long-term follow-up data of 648 hepatitis D patients recruited by 14 centres in 11 countries. Liver-related clinical endpoints were defined as hepatic decompensation (ascites, encephalopathy and variceal bleeding), liver transplantation, hepatocellular carcinoma or liver-related death. RESULTS:Patient data were available from all continents but Africa: 22% from Eastern Mediterranean, 32% from Eastern Europe and Central Asia, 13% from Central and Southern Europe, 14% from South Asia (mainly Pakistan) and 19% from South America (mainly Brazil). The mean follow-up was 6.4 (.6-28) years. During follow-up, 195 patients (32%) developed a liver-related clinical event after 3.5 (±3.3) years. Liver cirrhosis at baseline and a detectable HDV RNA test during follow-up were associated with a worse clinical outcome in multivariate regression analysis while patients receiving interferon alfa-based therapies developed clinical endpoints less frequently. Patients from South Asia developed endpoints earlier and had the highest mortality. CONCLUSIONS:The HDIN registry confirms the severity of hepatitis D and provides further evidence for HDV viraemia as a main risk factor for disease progression. Hepatitis D seems to take a particularly severe course in patients born in Pakistan. There is an urgent need to extend access to antiviral therapies and to provide appropriate education about HDV infection.