Detection of the hepatitis B surface antigen in patients with occult hepatitis B using an assay with enhanced sensitivity

Patients with occult hepatitis B infection (OBI) have undetectable hepatitis B surface antigen (HBsAg) by conventional assays but detectable hepatitis B virus (HBV) DNA in blood/liver. We evaluated the key performance characteristics of a sensitive HBsAg assay (ARCHITECT HBsAg Next Qualitative Assay, referred as NEXT) with respect to HBsAg detection. Assay precision, sample carryover and seroconversion sensitivity of NEXT were evaluated. HBsAg was measured by NEXT in 1,138 individuals, including 1,038 patients who attended liver clinics in a tertiary hospital (200 HBV DNA-positive blood donors whose HBsAg was undetectable by conventional assays, and 38 patients receiving immunosuppressive therapy, 800 chronic hepatitis B patients with HBsAg seroclearance) and 100 HBsAg-negative subjects recruited from a community project. The within-run and within-laboratory coefficients of variation were <6% for the positive sample pools. In 9 seroconversion panels tested, NEXT allowed an earlier HBsAg detection than conventional assays. NEXT detected HBsAg in 10/200 (5%) HBsAg-negative blood donors, 1/20 (5%) and 0/18 HBsAg-negative patients with and without HBV reactivation respectively, and 59/800 (7.3%) patients with HBsAg seroclearance. HBsAg was detectable by NEXT in 27.8%, 8.2%, 6.9%, 3.8% and 1.9% samples at <3, 3–5, >5–8, >8–11, and >11 years after HBsAg seroclearance, respectively. Seven out of 100 HBsAg-negative community identified subjects was tested positive by NEXT. Comparing with conventional HBsAg assays, NEXT demonstrated a higher sensitivity and conferred an increment of 5–7% detection rate in patients with OBI, thereby helping in identifying HBV carriers and prevention of OBI-associated HBV transmission and reactivation.

High Frequency Occult Hepatitis B Virus Infection Detected in Non-Resolved Donations Suggests the Requirement of Anti-HBc Test in Blood Donors in Southern China

BackgroundMost Chinese Blood Centers adopted mini pool (MP) nucleic acid testing (NAT) for HBV screening due to high cost of Individual donation (ID) NAT, and different proportions of MP-reactive but ID-non-reactive donations (MP+/ID−, defined as non-resolved donations) have been observed during daily donor screening process. Some of these non-resolved donations are occult HBV infections (OBIs), which pose potential risk of HBV transmission if they are not deferred. This study is aimed to further analyze these non-resolved donations.MethodsThe non-resolved plasma samples were further analyzed by serological tests and various HBV DNA amplification assays including quantitative PCR (qPCR) and nested PCR amplifying the basic core and pre-core promoter regions (BCP/PC; 295 base pairs) and HBsAg (S) region (496 base pairs). Molecular characterizations of HBV DNA+ non-resolved samples were determined by sequencing analysis.ResultsOf 17,226 MPs from 103,356 seronegative blood donations, 98 MPs were detected reactive for HBV. Fifty-six out of these 98 (57.1%) reactive MPs were resolved as HBV DNA+, but the remaining 42 pools (42.9%, 252 donations) were left non-resolved with a high rate (53.2%) of anti-HBc+. Surprisingly, among 42 non-resolved MPs, 17 contained one donation identified as OBIs by alternative NAT assays. Sequence analysis on HBV DNAs extracted from these OBI donations showed some key mutations in the S region that may lead to failure in HBsAg detection and vaccine escape.ConclusionA total of 53.2% of the non-resolved donations were anti-HBc+, and OBIs were identified in 40.5% of these non-resolved pools. Therefore, non-resolved donations with anti-HBc+ might pose potential risk for HBV transmission. Our present analysis indicates that anti-HBc testing in non-resolved donations should be used to identify OBIs in order to further increase blood safety in China.

Novel Biomarkers of Hepatitis B Virus and Their Use in Chronic Hepatitis B Patient Management

Even though an approved vaccine for hepatitis B virus (HBV) is available and widely used, over 257 million individuals worldwide are living with chronic hepatitis B (CHB) who require monitoring of treatment response, viral activity, and disease progression to reduce their risk of HBV-related liver disease. There is currently a lack of predictive markers to guide clinical management and to allow treatment cessation with reduced risk of viral reactivation. Novel HBV biomarkers are in development in an effort to improve the management of people living with CHB, to predict disease outcomes of CHB, and further understand the natural history of HBV. This review focuses on novel HBV biomarkers and their use in the clinical setting, including the description of and methodology for quantification of serum HBV RNA, hepatitis B core-related antigen (HBcrAg), quantitative hepatitis B surface antigen (qHBsAg), including ultrasensitive HBsAg detection, quantitative anti-hepatitis B core antigen (qAHBc), and detection of HBV nucleic acid-related antigen (HBV-NRAg). The utility of these biomarkers in treatment-naïve and treated CHB patients in several clinical situations is further discussed. Novel HBV biomarkers have been observed to provide critical clinical information and show promise for improving patient management and our understanding of the natural history of HBV.

Molecular surveillance of HIV, HBV, and HCV amongst blood donors in five Chinese regions

Hepatitis B (HBV), hepatitis C (HCV) and human immunodeficiency virus (HIV) are transfusion transmissible infections (TTIs) agents that threaten the safety of the blood supply. Surveillance of the variance of those viruses is an important way to monitor their diversity and evolution to improve safety in blood transfusion. In this study, we characterized the specimens of blood donors from 13 blood centers located in 5 Chinese regions.Samples collected between 2014 and 2017 were screened with serological and molecular tests conducted on Abbott ARCHITECT and m2000 platforms. Sequencing was used to determine the classifications. The HBV immune escape mutations were also analyzed for assessing vaccine breakthrough risks and challenges for diagnostic tests. For HIV, 11 genotypes or recombinants were identified. The predominant genotype was C, which accounts for 42%. For HBV, the genotypes of B, C and D were identified, with B and C predominating. The major subgenotype was B2, comprising 84.1% of all infections. 79 out of 113 (69.9%) samples carried escape mutations in the “a” determinant region with 69 (87.3%) multiple mutants and 15 (19%) escape mutants which will affect HBsAg detection. For HCV, 7 genotypes or subtypes were identified. The major genotype was 1b (48%), followed by 6a (16.7%) and 2a & 3a (10%). This study provides the information of diversity of HBV, HCV and HIV strains circulating in blood centers from 5 regions in China. These data can also be scientific basis for development of detection assays that mitigate the impact of viral diversity on performance.ImportanceThe prevalence of TTIs in blood donations is important for evaluating blood safety and it can also reflect the burden of these disease among populations. Virus variance is threat to blood safety due to it may affect assays detection by nucleic acid, antigen and antibody-based methods in blood donors. HIV, HBV and HCV exhibit high degrees of genetic diversity, with different strains predominating in different geographic locations. The aim of this study is to assess the diversity of HBV, HCV and HIV among blood donors in China. In this study, 13 blood centers located in 5 Chinese regions were involved and the most informative phylogenetic regions of each virus had been sequenced. This will benefit for viral monitoring by subtype/genotype analyses to determine whether the distributions of variants are changing over time and geographically, and to speculate whether previously rare subtypes are becoming established in blood donors in China.

Chronic hepatitis b associated without/with a delta agent in Kyrgyzstan (epidemiological situation, clinical features)

Objective. To compare epidemiological, clinical and laboratory characteristics of chronic hepatitis B (ChHB) associated with/without delta agent (ChHB+DV) study.Materials and methods. The Kyrgyzstan State Reporting Form No. 12 covering 2010–2017 period was examined. For this, 133 and 130 case histories of ChHB and ChHB+DV patients, respectively, were analyzed. The data were statiastically processed by using Microsoft Office Excel software.Results and discussion. Over the 2010–2017 period, prevalence of the “HBV Carrier” (60.4 ) was higher by 20-fold than that one for ChHB [3.8 , 95% CI (2.4–4.0)] and CVHD [3.4 , 95% CI (2.2–3.4)], as the vast majority of patients were not thoroughly examined after detecting HBsAg, and the HBV Carrier was empirically diagnosed at the primary health care units. As a result, routine case definitions for such conditions were revised and an improved system of epidemiological surveillance of viral hepatitis was developed, according to the 2016 WHO recommendations approved by the Ministry of Health of the Kyrgyz Republic (Order No. 524, dated of July 20, 2018). Asthenia was observed in ~60% of patients in both groups, whereas arthralgia — in ~5–10% of patients, more often in those comorbid with ChHB+DV, and myalgia — in as low as ~3% of cases. Impaired central nervous system functions manifested as headache and restless sleep were evenly recorded in about 10–15% of patients, without significant difference between groups. In contrast, dominating dyspeptic manifestations such as poor appetite (72±3.9% vs. 20.6±3.5%, p < 0,05), nausea (23.8±3.7% vs. 7.3±2.3%, p < 0,05), vomiting (12.3±2.6% vs. 3.3±1.5%, p < 0,05) and flatulence (27±3.9% and 13±2.9%, p < 0,05) were revealed in ChHB+DV patients. Pain in the right hypochondrium was noted in 52–56% of patients, insignificantly differed between patient groups. Incidence of yellowness of the sclera and skin layers as well as skin itching were recorded by 2–3 and 8 times, respectively, more frequently in ChHB+DV patients. A more profound cytolysis and signs of altered bilirubin metabolism were also more common in HBV patients comorbid with the delta agent. Thus, a more severe ChHB+DV course requires that all patients with primary HBsAg detection were mandatorily examined for anti-HDV antibodies to ensure early diagnostics and timely organization of the secondary and tertiary preventive measures in the Kyrgyzstan.

Prenatal care: missed opportunity for HBV prevention in women of childbearing age in rural Senegal

Background Perinatal transmission of hepatitis B virus (HBV) constitutes an important risk in highly endemic countries including Senegal. Although the prevalence of chronic HBV infection is estimated at 11% in this country, specific data on women of childbearing age are sorely lacking. We described in this study the prevalence of the HBV antigen (HbsAg) in women of childbearing age in rural Senegal, as well as general knowledge on HBV and hepatitis B status awareness. Methods A cross-sectional study including HBV screening was conducted at home in the rural Niakhar area. Chronic HBV infection was determined through HBsAg detection using dried blood spots. Socio-demographic and behavioral data were collected through standardized face-to-face questionnaires. The analyzes included 368 women aged 15-49 (67% married; 65% with at least 1 child) enrolled from October 2018 to March 2019. Results Preliminary results show that 49 women (13%) had positive HBsAg. Only 68 women (18%) have already heard about HBV. Among them, 53% knew that there exists an HBV vaccine and 78%, 75% and 67% correctly answered that HBV can be transmitted through blood contact, childbirth and sexual intercourse, respectively. Among the 233 (63%) women who had already given birth, 76% had attended at least 4 antenatal care sessions and 74% had given birth in a healthcare facility for their last pregnancy. However, only 1% reported to have already been screened for HBV. Main reasons reported for not having been screened were having never heard about this test (80%) and not having been offered screening during antenatal care sessions (10%). Conclusions General knowledge and awareness of HBV status are particularly low in women of childbearing age living in rural Senegal, despite high antenatal care sessions attendance. Given the high prevalence of chronic HBV infection found in this population, it is urgent to ensure systematic HVB screening and to provide adequate information to women during pregnancy. Key messages The prevalence of chronic HBV is high in women of childbearing age in Senegal. National recommendations for women attending antenatal care sessions should include HBV screening and counselling.