Background HIV-HBV coinfection accelerates liver disease, yet the immunological mechanisms underlying adverse outcomes remain incompletely characterized in African populations. We investigated relationships between HBV reactivation, cytokine dysregulation, T-cell dysfunction, and disease progression in a Nigerian cohort. Methods We screened 1,139 participants across four Nigerian states. Of 344 HIV-positive individuals, 53 (15.4%) had HBV coinfection. For detailed immunological and longitudinal analyses, 59 coinfected participants with complete datasets were included in the mechanistic cohort. Comprehensive assessments including HBV DNA quantification, S-gene sequencing, cytokine profiling, and HBV-specific T-cell responses were performed on 59 coinfected patients with longitudinal follow-up. Results Phylogenetic analysis indicated 71.4% (30/42) of cases with rising HBV DNA were consistent with reactivation. HBV genotype E predominated (94.3%). Coinfected patients demonstrated elevated IL-6 and TNF-α with reduced IFN-γ compared with HIV-monoinfected controls (all p < 0.001). The IL-6/IFN-γ ratio correlated with HBV viral load (r = 0.74), APRI score (r = 0.71), and CD4+ count (r = −0.64; all p < 0.001). HBV-specific polyfunctional CD8+ T-cells were markedly reduced (median 0.08% vs 3.8% in controls; p < 0.001). In multivariable Cox regression, IL-6/IFN-γ ratio >4.0 (HR 4.12, 95% CI 1.86–9.14), CD4+ <200 cells/µL (HR 3.24, 95% CI 1.58–6.64), and APRI >1.0 (HR 2.86, 95% CI 1.34–6.11) independently predicted progression, whilst preserved T-cell polyfunctionality was protective (HR 0.32, 95% CI 0.15–0.68). Conclusions HIV-HBV coinfection was characterized by HBV reactivation, cytokine imbalance, and T-cell exhaustion, which were associated with disease progression and may inform risk stratification.

Background/Objectives: Chronic hepatitis B (CHB) and type 2 diabetes mellitus (T2DM) frequently coexist. This study aimed to investigate the impact of T2DM and glycemic control on antiviral efficacy in CHB patients. Methods: This single-center, retrospective cohort study included treatment-naïve CHB patients who initiated nucleos(t)ide analogue (NA) therapy between January 2019 and January 2024. The primary endpoint was a complete virological response (CVR), defined as achieving HBV DNA levels below 20 IU/mL after 48 weeks of treatment. Results: The CHB + T2DM group (n = 81) demonstrated a significantly lower CVR rate than the CHB group (n = 106) (26.0% vs. 41.2%, p = 0.038). Multivariate analysis identified T2DM as an independent negative predictor of a CVR (OR = 0.400, 95% CI: 0.196–0.815, p = 0.012). Within the CHB + T2DM subgroup, adequate glycemic control (HbA1c < 7%) was associated with a higher CVR (38.7% vs. 16.7%, p = 0.034). Patients newly diagnosed with diabetes at enrollment showed a higher rate of HBeAg loss than those with pre-existing diabetes (57.1% vs. 10.0%, p = 0.036). Regarding antiviral regimens, entecavir-treated CHB + T2DM patients had a lower CVR than CHB controls (18.8% vs. 46.2%, p = 0.015). Furthermore, tenofovir-based regimens showed a more favorable antiviral trend than entecavir in CHB patients with T2DM. Conclusions: Comorbid T2DM was an independent risk factor for impaired antiviral efficacy in CHB patients. Optimal glycemic control may improve virological outcomes. These findings suggest that the early diagnosis and management of T2DM could enhance antiviral treatment efficacy in CHB patients.

The rising prevalence of chronic hepatitis B (CHB) and metabolic dysfunction-associated fatty liver disease (MAFLD) comorbidity necessitates a deeper understanding of their clinical interplay. Although miR-122 is a known liver-specific regulator of chronic liver diseases, its specific role in the context of CHB-MAFLD remains elucidated. In this study, serum analysis revealed that CHB-MAFLD patients exhibit significantly lower levels of HBV DNA, pgRNA, and HBsAg compared to patients with CHB alone, which correlated with elevated miR-122 expression. Through bioinformatics and molecular assays, SOX4 was identified as a direct downstream target of miR-122-5p. Utilizing an HBV-infected steatotic cell model, we observed that the comorbid state suppresses HBV replication markers-including DNA, pgRNA, HBsAg, and HBcrAg-while concurrently upregulating the miR-122-5p/SOX4 axis. Functional experiments demonstrated that miR-122-5p and SOX4 both act as inhibitors of HBV activity, as their knockdown enhanced viral replication, while their overexpression led to significant suppression. These findings suggest that the metabolic environment in CHB-MAFLD may naturally suppress HBV replication through the activation of a novel miR-122/SOX4 regulatory axis. This study highlights miR-122 as a potential therapeutic target and provides a mechanistic basis for the altered viral kinetics observed in patients with dual liver pathology. Trial Registration: Chinese Clinical Trial Registry identifier: ChiCTR2200063555.

To analyze the clinical and hepatic pathological characteristics of patients with chronic hepatitis B (CHB) alone or with non-alcoholic fatty liver disease (NAFLD), who have normal alanine aminotransferase (ALT) levels. The patients with normal ALT levels and pathologically diagnosed CHB alone or in combination with NAFLD were enrolled, the demographic, laboratory, and pathological data were collected and analyzed. Among 391 enrolled CHB patients with normal ALT levels, 107 individuals combined with NAFLD. The incidence of significant liver injury (G and/or S ≥ 2) in patients with CHB alone was lower significantly than that of patients with CHB and NAFLD (64.08% vs. 78.50%, P < 0.05), especially by the subgroup analyses in HBeAg positive, male, age > 30 years (P < 0.05). In all patients, who with negative HBcAg expression had lower HBV DNA levels than those with positive HBcAg expression (P < 0.001). Patients with G and/or S ≥ 2 had a higher proportion of HBcAg cytoplasm/cytoplasmic nucleus expression type compared to patients with liver tissue G and S < 2 (32.42% vs. 16.67% in CHB alone, 36.91% vs. 21.74% in CHB + NAFLD, all P < 0.001). Aspartate aminotransferase to ALT ratio index (AAR) demonstrated relatively superior efficacy in diagnosing significant liver fibrosis among patients with CHB and NAFLD, while fibrosis-4 (FIB-4) exhibited superior performance in patients with CHB alone. Patients with normal ALT can still demonstrate significant liver tissue damage. The combination of NAFLD will significantly increase the incidence of significant liver injury in patients with CHB, especially in those who are HBeAg positive, male, over 30 years old.

Background and AimsNovel antiviral approaches capable of permanently inactivating the intrahepatic HBV DNA reservoir, the covalently closed circular DNA (cccDNA) and HBV DNA integrated into the host genome, are urgently needed. This study evaluated adenine base editing as a strategy to disrupt HBV replication by introducing mutations in the overlapping HBs/polymerase open reading frame (ORF).MethodsAn adenine base editor (ABE) and 3 guide RNAs (gS1-gS3) were designed to introduce missense mutations within the HBs/polymerase ORF. ABE mRNA and individual gRNAs were co-transfected into HBV-infected HepG2-hNTCP cells and primary human hepatocytes. Antiviral efficacy was further assessed in HepG2.2.15 and PLC/PRF/5 cells harboring integrated HBV DNA. In vivo, lipid nanoparticles (LNP)-mediated delivery of ABE mRNA and gRNAs was evaluated in HBVcircle DNA-transduced mice and in HBV-infected human liver-chimeric mice. The impact of HBs editing on hepatitis D virus (HDV) release was assessed using PLC/PRF/5 and Huh7 cell-based HDV replication models.ResultsAdenine base editing efficiently reduced HBsAg production and HBV replication in vitro by targeting both cccDNA and integrated HBV DNA. A single LNP injection of ABE-gS2 resulted in undetectable HBsAg in HBVcircle mice, while two injections achieved a 90% reduction in serum HBsAg in HBV-infected human liver chimeric mice. HBV DNA replication was also inhibited in vivo. Furthermore, HBs ORF base editing markedly suppressed HDV release in vitro.ConclusionsAdenine base editing of the HBs ORF effectively impairs HBV replication and HBsAg production in vitro and in vivo and concomitantly inhibits HDV release, highlighting its therapeutic potential.

Chronic hepatitis B (CHB) with positive HBeAg status constitutes a significant contributor to the development of liver cirrhosis and hepatocellular carcinoma. Pegylated interferon-alpha (pegIFNα) is a common treatment, but its response rate remains limited, and the underlying mechanisms are not fully understood. Eighty-two HBeAg-positive CHB patients were enrolled. miR-194-5p expression, HBeAg, and HBV DNA levels were detected using qRT-PCR, ELISA, and quantitative PCR, respectively. ROC and logistic regression analyses were performed. Cellular experiments, including dual-luciferase reporter and rescue assays, along with Western blot analysis of JAK-STAT pathway proteins, were conducted to verify targeting and function. Complete response (CR) patients had significantly lower baseline HBV DNA than suboptimal response (SR) patients. After 48 weeks of pegIFNα therapy, miR-194-5p expression decreased notably in the CR group and correlated positively with HBV DNA and HBeAg levels. miR-194-5p predicted treatment response with an AUC of 0.831 and was an independent predictor. Mechanistically, miR-194-5p targeted SOCS2. Functional studies demonstrated that miR-194-5p overexpression enhanced, while SOCS2 supplementation attenuated, pegIFNα-induced phosphorylation of STAT1/STAT2, thereby influencing cell viability and inflammatory factor expression (TNF-α, IL-6, IL-1β). miR-194-5p may predict pegIFNα response in HBeAg-positive CHB. It regulates interferon signaling by targeting SOCS2 and modulating the JAK-STAT pathway activation, suggesting the miR-194-5p/SOCS2 axis as a potential therapeutic target.

Nucleos(t)ide analog (NAs) treatment achieves limited HBsAg loss rates; switching to or adding Peginterferon (PegIFN) therapy may improve outcomes. This study aimed to evaluate the efficacy of adding PegIFNα-2b therapy in chronic hepatitis B (CHB) patients who are virally suppressed by NAs. 250 CHB patients on NAs treatment with HBsAg < 1500 IU/mL, serum HBV DNA < 20 IU/mL, and ALT ≤ 1.5 × ULN were enrolled. Patients continued their NAs regimen and initiated PegIFNα-2b (180µg/week) for 48 weeks, with follow-up until week 72. In addition to routine biochemistry, HBsAg and HBV RNA levels were measured at each visit; HBcrAg was assessed at baseline, week 12, and week 24. 214 patients completed 48 weeks of PegIFNα-2b therapy, and 34.6% achieved HBsAg loss at week 72. Lower baseline HBsAg was associated with higher rates of HBsAg clearance. Specifically, patients with baseline HBsAg < 10 IU/mL and 10-100 IU/mL achieved HBsAg clearance rates of 76.2% and 45.5% at week 72, respectively. In univariate analysis, baseline HBsAg and its decline at week 12 demonstrated the strongest predictive performance for HBsAg loss, while younger age and higher ALT were also associated with HBsAg clearance. However, in multivariate analysis, only age and HBsAg decline at week 12 remained independent predictors of HBsAg loss. CHB patients with low HBsAg levels and virologic suppression on NAs can achieve significant HBsAg loss after adding PegIFNα-2b. A two-step strategy based on baseline HBsAg and week-12 HBsAg decline may aid patient selection and treatment optimization.

Low-level viremia increases the risk of adverse long-term outcomes in entecavir-treated patients with chronic hepatitis B.

Simultaneous and ultrasensitive detection of HBV DNA and RNA in CHB by utilizing Cas12a/Cas13a-based dual-target strategy

This study aimed to analyze the risk factors and predicted model for clinical relapse after discontinuation of antiviral therapy in patients with chronic hepatitis B (CHB). A retrospective analysis was conducted on the clinical data of 99 CHB patients who met the discontinuation criteria and were treated at Southern Central Hospital of Yunnan Province (The First People's Hospital of Honghe State) from March 2020 to December 2022. All subjects received nucleos(t)ide analogs (NAs) or interferon-based antiviral therapy and discontinued treatment once they met the cessation criteria, followed by a 2-year follow-up. Based on relapse status, patients were divided into a relapse group and a non-relapse group. Clinical characteristics were compared between the two groups. A multivariate logistic regression analysis was performed to analyze the independent risk factors for clinical relapse within 2years after treatment cessation. During the 2-year follow-up, 45 patients (45.45%) experienced clinical relapse after discontinuation. Compared with the non-relapse group, the relapse group exhibited significantly higher age, HBsAg levels at treatment cessation, and HBV DNA load at discontinuation (p < 0.05), as well as a shorter total duration of antiviral therapy (p < 0.05). Multivariate analysis revealed that age, total antiviral treatment duration, HBV DNA load at discontinuation, and HBsAg levels at cessation were independent risk factors for clinical relapse of CHB patients (p < 0.05). A combined predictive model was constructed based on multivariate logistic regression coefficients: combined model = -17.497 + 0.181 × age + (-0.123) × total antiviral duration + 1.746 × HBV DNA at discontinuation + 0.032 × HBsAg at discontinuation. ROC analysis demonstrated that the AUC of the combined model was 0.945 (95% CI: 0.902-0.987) for predicting 2-year clinical relapse, with a sensitivity of 91.11% and specificity of 83.33%. Spearman correlation analysis indicated that patient age and HBV DNA load at discontinuation were negatively correlated with time to relapse (p < 0.05), whereas HBsAg levels showed no significant correlation with total antiviral duration (p > 0.05). Age, HBV DNA load at discontinuation, HBsAg quantification at discontinuation, and the total antiviral duration were identified as key factors influencing clinical relapse after cessation of antiviral therapy in patients with CHB. A predictive model incorporating these factors demonstrated good clinical predictive value.

Comparison of serum HBcrAg, HBV RNA, and high-sensitivity HBV DNA as predictors of efficacy in patients with chronic hepatitis B treated with entecavir

BACKGROUNDHepatitis B Virus (HBV) remains a global public health challenge, affecting over 296 million people, many of whom are asymptomatic. Incidentally diagnosed carriers provide a critical window for early intervention and prevention. Understanding their hematological profile and viral burden can help inform risk assessment and clinical management.AIMTo evaluate hematological parameters and HBV viral load in incidentally detected asymptomatic hepatitis B surface antigen positive patients during routine health screenings.METHODSA cross-sectional observational study was conducted from June 2024 to March 2025 at Dr. D. Y. Patil Medical College, Hospital and Research Centre a tertiary care hospital in Pune, Maharashtra, India, involving 100 hepatitis B surface antigen-positive patients and 20 healthy controls. Hematological and liver function parameters were assessed, and quantitative HBV DNA analysis was performed using real-time polymerase chain reaction. Statistical analysis was conducted using GraphPad Prism v8.0.RESULTSMarked variations were detected in hemoglobin levels (P < 0.0001), percentage of neutrophils (P = 0.0006), percentage of lymphocytes (P = 0.0031), serum glutamic-oxaloacetic transaminase activity (P = 0.0013), and alanine aminotransferase levels (P = 0.0001) when comparing HBV-infected individuals with the control group. Conversely, differences in total leukocyte count, percentage of monocytes, percentage of eosinophils, platelet count, total bilirubin, and serum glutamic-pyruvic transaminase values were not statistically significant. Viral load > 2000 IU/mL was found in 17 patients, and < 2000 IU/mL in 24 patients. Viral load positively correlated with conjugated bilirubin, serum glutamic-oxaloacetic transaminase, serum glutamic-pyruvic transaminase, and alkaline phosphatase, and negatively with albumin and the albumin-globulin ratio.CONCLUSIONIncidentally detected HBV infections present an opportunity for early disease detection. Hematological and viral markers can guide clinical decisions. Routine screening and contact tracing are essential strategies to control HBV transmission and progression.

Objective: To explore the antiviral efficacy and safety profile of nucleos(t)ide analogue (NA) mono-or combination therapy with interferon-α (IFN-α) in children 1-6 years old with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB). Methods: Seventy-eight children 1-6 years old with HBeAg-positive CHB from multiple centers from November 2020 to August 2023 were enrolled as research subjects and were divided into the NA group (n=24) and the NA+IFN group (n=54) according to the antiviral treatment regimen. Oral lamivudine or entecavir and interferon-α (IFN-α) or pegylated interferon-α (PEG-IFN-α) were administered to the NA and NA+IFN groups. The HBV DNA negativity rate, HBeAg negative seroconversion rate, and HBsAg clearance rate were compared at 48 weeks between the two groups, with further stratification by 1-3 and 4-6 years old for the aforementioned comparison. An independent samples t-test or Mann-Whitney U test was used for inter-group comparison of quantitative data. The χ2 test or Fisher's exact test was used for inter-group comparison of categorical data. The Benjamini-Hochberg false discovery rate (FDR) method was used for multiple comparison correction. Results: The HBV DNA negativity rate [81.82% (27/33) vs. 41.18% (7/17), P=0.004, q=0.018] and HBsAg clearance rate [36.36% (12/33) vs. 0 (0/17), P=0.004,q=0.018] were higher in the NA+IFN group than in those in the NA group at 48 weeks among children 4-6 years old (n=50), with statistically significant differences. However, there were no statistically significant differences in the HBV DNA negativity rate [57.14% (12/21) vs. 71.43% (5/7)], HBeAg negative seroconversion rate [47.62% (10/21) vs. 28.57% (2/7)], and HBsAg clearance rate [19.05%(4/21) vs. 28.57%(2/7)] for children 1-3 years old (n=28) between the NA+IFN group and the NA group. Conclusion: The antiviral efficacy of the NA+IFN combination therapy group was superior to that of the NA monotherapy group at 48 weeks in children with CHB 4-6 years old. However, there was no statistically significant difference in the efficacy of the two antiviral regimens among young children 1-3 years old, hinting that NA therapy may be a priority for young children.

Limited research exists on immune checkpoint inhibitors (ICIs)' antiviral efficacy in HBV-related hepatocellular carcinoma (HCC). This study aimed to evaluate the impact of ICIs on multiple HBV markers and its correlation with HCC prognosis. A retrospective cohort study was performed on 318 HBV-related HCC patients, including 268 who received ICIs (with/without targeted therapy; ICI group) and 50 who received targeted therapy alone (non-ICI group). Levels of quantitative HBsAg (qHBsAg), HBV DNA, HBV RNA and HBcrAg were monitored. Tumor response was evaluated using RECIST 1.1. Over a median 8.1-month follow-up, the ICI group showed a significantly greater decrease in all HBV markers. At week 96, the ICI group showed significantly higher cumulative incidences of HBsAg response (loss or ≥0.5 Log10 decline) (41.6% vs. 14.9%, p=0.006) and HBcrAg response (negativity or ≥1 Log10 decline) (46.3% vs. 18.1%, p=0.007) than non-ICI group, and a significantly faster achievement rate of HBV RNA response (negativity or ≥0.5 Log10 decline) (HR: 1.80, 95% CI: 1.08-2.99, p=0.021). Notably, the PD-1 inhibitors showed significantly better virological response efficacy than PD-L1 inhibitors (HR=2.04-5.43). The patients with lower levels of HBV markers at enrollment demonstrated significantly better overall survival (OS). Moreover, patients achieving virological response were associated with significantly better survival outcomes and tumor response, with HR ranging 1.64-8.06. ICIs demonstrate dual therapeutic effects in HBV-related HCC, significantly reducing multiple HBV markers while concurrently enhancing prognosis in combination with antiviral therapy, emphasizing the importance of sustained virological suppression for optimal HCC outcomes.

BackgroundHepatitis B virus (HBV) remains a major public health challenge in sub-Saharan Africa. Somalia, with its fragile healthcare system and high-risk population, lacks up to date data on HBV epidemiology. This study assessed HBV prevalence, associated risk factors, and viral load among hospital patients in Mogadishu, Somalia.MethodsA cross-sectional survey was conducted in four major hospitals in Mogadishu. Socio-demographic and clinical risk data were collected using structured questionnaires. Blood samples from 270 participants were screened for HBsAg using a rapid test, with positives confirmed and quantified for HBV DNA by real-time PCR. Bivariate and multivariate analyses identified independent risk factors.ResultsHBsAg prevalence was 11.5% (n = 31), but only 2.2% of all participants (6/270; 19.4% of HBsAg-positives) had detectable HBV DNA. The strongest independent predictors were family history of HBV (AOR: 5.08, 95% CI 1.90–13.60) and history of tooth extraction (AOR: 3.18, 95% CI 1.24–8.12). Socio-economic factors, facility variations, and low vaccination were also observed. Classical behavioral risk factors were infrequent and not significant. Among PCR positive cases, only one presented with a high viral load (>20,000 IU/mL).ConclusionHBV poses a substantial, ongoing threat to the Somali population, especially in hospital-based settings. Household transmission and unsafe medical procedures are the most significant risk factors. Improving infection control, expanding vaccination, and prioritizing molecular diagnostics will be critical to effective HBV control in Somalia.

BackgroundBoesenbergia rotunda (fingerroot) is widely used in traditional medicine, and its bioactive compound panduratin A has demonstrated potent antiviral properties. However, the mechanistic basis underlying its anti-hepatitis B virus (HBV) activity remains to be fully elucidated.MethodsHBV-infected human hepatocytes (imHCs) were treated with B. rotunda extract, panduratin A, or pinostrobin. Intracellular HBV DNA, secreted HBsAg and HBeAg, and pregenomic RNA (pgRNA) were quantified in dose- and time-dependent experiments. Luciferase reporter assays were used to assess HBV promoter activity. The roles of HNF1α and HNF4α were evaluated by siRNA-mediated knockdown and ectopic gene expression. Drug interaction studies were performed using the KDM5 inhibitor GS-5801 and the capsid assembly modulator NVR-3778. A 3D liver spheroid model was used to validate antiviral effects on HBV DNA and cccDNA. Gene interaction network analysis was conducted to identify central regulatory pathways.ResultsB. rotunda extract, panduratin A, and pinostrobin significantly suppressed intracellular HBV DNA, HBsAg, HBeAg, and pgRNA. Panduratin A exhibited the strongest antiviral activity and inhibited preS1, preS2, and core promoter activities. Panduratin A markedly downregulated HNF1α expression, with only modest effects on HNF4α. Knockdown of HNF1α significantly reduced the antiviral efficacy of panduratin A, whereas ectopic HNF1α expression rescued its inhibitory effects. Co-treatment with GS-5801 produced synergistic activity, and combination with NVR-3778 yielded additive antiviral effects. In 3D liver spheroids, panduratin A reduced intracellular HBV DNA and cccDNA with minimal cytotoxicity. Network analysis further identified HNF1α as a key regulatory node modulated by panduratin A.ConclusionPanduratin A is a potent anti-HBV compound that acts primarily through HNF1α-dependent suppression of HBV transcription and replication. Its efficacy in combination therapy and in 3D liver models highlights its potential as a promising candidate for future HBV treatment strategies.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1186/s13020-025-01285-w.

This multicenter study evaluated serum HBV RNA as a prognostic biomarker for nucleos(t)ide analogue (NA) therapy in chronic hepatitis B (CHB), focusing on correlations with HBV DNA/HBsAg and predictive value for virological response (VR) and HBeAg seroconversion. From 2020 to 2024, 427 treatment-naive HBeAg-positive CHB patients across nine Chinese hospitals were enrolled. Longitudinal HBV RNA-DNA-HBsAg correlations were analyzed during 24-month NA therapy. Predictive performance for VR at 12 months and HBeAg seroconversion at 24 months was assessed. Non-VR patients had higher baseline HBV DNA (median 7.42 vs. 5.38 log10 IU/mL, P = 0.001) and HBV RNA (6.63 vs. 4.60 log10 copies/mL, P = 0.001). Pretreatment HBV RNA correlated with HBV DNA (r = 0.638, P = 0.001), shifting to HBsAg post-24-month treatment (r = 0.917, P = 0.0005). HBV DNA and RNA levels at 6 months independently predicted therapeutic outcomes. For VR, HBV DNA demonstrated a hazard ratio (HR) of 0.16 (P < 0.001) with an area under the ROC curve (AUROC) of 0.889, while HBV RNA showed an HR of 0.77 (P = 0.03) and AUROC of 0.754. For HBeAg seroconversion, HBV DNA yielded an HR of 0.01 (P = 0.015) with AUROC 0.900, and HBV RNA exhibited an HR of 0.72 (P = 0.03) and AUROC 0.703, underscoring their consistent prognostic value at this critical timepoint. HBV RNA dynamics transition from DNA- to HBsAg-associated correlations during NA therapy. Early declines in HBV DNA/RNA at 6 months predict VR and HBeAg seroconversion, establishing this time as a a robust and clinically practical early prediction point. HBV RNA complements existing biomarkers for optimizing CHB management.

Investigation on CRISPR-Cas12a-split crRNA system for successively detecting DNA and RNA in one tube.

Occult hepatitis B virus infection (OBI) refers to the detection of HBV DNA in blood or liver tissue in individuals who test negative for hepatitis B surface antigen (HBsAg), excluding the early phase of acute HBV infection. This hidden form of infection represents a substantial but frequently underestimated risk to blood transfusion safety, particularly in settings with limited resources. In Ethiopia and other low-income countries, routine screening of blood donors often depends solely on HBsAg tests, which may overlook symptom-free carriers with low viral loads. This study was conducted to estimate the prevalence of OBI among HBsAg-negative blood donors and to assess its potential contribution to undetected HBV transmission in central Ethiopia. Five hundred and eighty-two plasma samples from HBsAg-negative donors were analyzed. Initial screening for anti-Hepatitis B core antibodies (anti-HBc) was carried out using an enzyme-linked immunosorbent assay (ELISA). Samples that tested positive for anti-HBc were then analyzed for HBV DNA using the Abbott m2000 automated real-time polymerase chain reaction (RT-PCR) system. At the same time, information on sociodemographic characteristics and potential clinical risk factors for HBV exposure was obtained through structured questionnaires. Data were analyzed with SPSS Version 21, and statistical significance was considered at a p value of less than 0.05. In this study, screening of 582 blood donors who were negative for HBsAg revealed that 135 (23.2%) donors, or nearly one-quarter, showed evidence of previous HBV exposure through anti-HBc reactivity. Subsequent molecular testing (PCR) identified a small proportion of these individuals carrying HBV DNA, confirming occult infection. Viral loads in all confirmed cases were low. Older donors, particularly those above 35 years of age, demonstrated a markedly higher rate of anti-HBc positivity. Overall, the occurrence of OBI was estimated at 1.7%, or approximately 17 cases for every 1000 blood donations, highlighting a critical gap in the current HBV screening protocol. The findings demonstrate that a notable number of blood donors in central Ethiopia carry occult HBV infection, equivalent to 17 cases per 1000 donations. This indicates a concealed yet important risk of HBV transmission through blood transfusion. These results underscore the need to enhance donor screening protocols by incorporating both anti-HBc testing and nucleic acid testing (NAT) to improve transfusion safety and prevent the spread of undetected HBV.

Suppression of HBV replication and expression by CRISPR/Cas9 ribonucleoproteins.