Biomarcadores convencionales y emergentes en hepatitis B / Conventional and emerging biomarkers in hepatitis B

A nivel mundial, 300 millones de personas están infectadas por el virus de la hepatitis B (VHB). A pesar de que existe una vacuna que previene la infección y se dispone de tratamiento antiviral que suprime la replicación del virus, no hay cura aún. El principal problema que evita la recuperación total del paciente, incluso para aquel que recibe tratamiento, es la persistencia de dos formas del genoma viral en los hepatocitos: el ADN circular covalentemente cerrado (ADNccc), el cual se encuentra en forma de episoma y tiene la capacidad de replicarse, y las secuencias lineales subge-nómicas que se integran en el genoma humano, con potencial oncogénico. Hasta el momento se dispone de unos pocos biomarcadores para monitorear o predecir la progresión de la enfermedad y la respuesta al tratamiento. Estos biomarcadores se detectan durante la infección, y son la base para la monitorización de la enfermedad y hacer un diagnóstico de la fase clínica de la infección. Recientemente han surgido nuevos biomarcadores como el antígeno relacionado con el core del virus de la hepatitis B (HBcrAg) y la detección del ARN del VHB, que parecen correlacionarse con los niveles transcripcionales del ADNccc, además, durante el tratamiento parecen ayudar a predecir la respuesta y podrían identificar aquellos a quienes se les puede suspender la terapia sin riesgo de recaída. En esta revisión, se describe la utilidad de los principales biomarcadores convencionales en hepatitis B, y se abordan los dos biomarcadores emergentes más estudiados que prometen evaluar el curso de la infección, al igual que determinar la progresión de la enfermedad y la respuesta al tratamiento.

Globally, 300 million people are infected with hepatitis B virus (HBV). Although there is a vaccine that prevents infection and antiviral treatment that suppresses the replication of the virus, there is still no cure. The main problem that prevents the total recovery of the patient, even for those who recei-ve treatment, is the persistence of two forms of the viral genome in hepatocytes: covalently close circular DNA (cccDNA), which is in the form of an episome that has the ability to replicate, and linear subgenomic sequences that are integrated into the human genome, with oncogenic potential. Few biomarkers are currently available to monitor or predict disease progression and response to treatment. These biomarkers are detected during infection and are the basis for monitoring the di-sease and making a diagnosis of the clinical phase of the infection. New biomarkers have recently emerged, such as hepatitis B core-related antigen (HBcrAg) and HBV RNA detection, which seem to correlate with cccDNA transcriptional levels while during treatment seem to help predict response, and could identify those for whom therapy can be discontinued without risk of relapse. In this review, the usefulness of the main conventional biomarkers in hepatitis B is described, and the two most studied emerging biomarkers are mentioned, which promise to evaluate the course of the infection, as well as to determine disease progression and treatment response.

Clinical study of hepatitis B vaccine in achieving hepatitis B surface antibody seroconversion in patients with functional cure

Abstract Hepatitis B Surface Antigen (HBsAg) seroclearance is the highest treatment goal recommended by the current guidelines for hepatitis B. Levels of antibodies to HBsAg (anti-HBs) are strongly associated with HBsAg recurrence, but hepatitis B vaccination may increase the anti-HBs seroconversion rate and reduce recurrence. We conducted a retrospective clinical study to ascertain the effect of this vaccination on the seroconversion rate and levels of protective anti-HBs after HBsAg. In this retrospective study, we distributed a questionnaire through an online survey platform to collect information related to hepatitis B vaccination in patients with functional cure of hepatitis B with Interferon-α (IFNα) therapy. We enrolled 320 patients who achieved functional cure from IFNα therapy. Of these, 219 patients had received hepatitis B vaccination according to their personal preference and drug accessibility after HBsAg seroclearance, whereas the remaining 101 patients did not receive hepatitis B vaccination. The anti-HBs seroconversion rate of 78.1% in the vaccinated group was significantly greater than that in the unvaccinated group (41.6%) (p < 0.001). Stratified comparisons with anti-HBs of ≥ 100 IU/L and ≥ 300 IU/L showed that both proportions in the vaccinated group were greater than those in the unvaccinated group (71.2% vs. 32.7% and 56.2% vs. 17.8%, respectively, all p-values < 0.001). Logistic regression analysis showed that the odds ratio of vaccination was 4.427, which was the strongest influencing factor for anti-HBs, reaching 100 IU/L or higher. Hepatitis B vaccination in patients after HBsAg seroclearance not only increased the anti-HBs seroconversion rate but also significantly increased antibody levels, with good safety, indicating the clinical value of vaccine therapy for patients with functional cure.

Initial screening and retest mode in apheresis platelet donors: a comparative analysis / 中国输血杂志

【Objective】 To investigate the effectiveness of current indicators in initial screening and retest before donation and access the optimal testing strategies. 【Methods】 Data of initial screening (rate method for ALT, colloidal gold method for HBsAg) and retest (rate method for ALT, ELISA for HBsAg) of 18 510 platelet donors in our center from January 2019 to December 2021 were collected, and the results were retrospectively analyzed and compared in terms of different years and number of donations. 【Results】 From 2019 to 2021, data of initial screening and retest of platelet donors were as follows: 1) the deferral rate of ALT and HBsAg was 12.98% (2 403/18 510) vs 0.26%(40/15 412); 2) the deferral rate of ALT was 13.19% (712/5 398) vs 0.20%(9/4 410)in 2019, 13.33% (873/6 549) vs 0.06%(3/5 387)in 2020 and 11.05% (725/6 563) vs 0.07%(4/5 615)in 2021; for initial screening, significant difference was noticed in ALT reactivity in 2021 as in comparison to other two years(P<0.05); 3) the reactive rate of HBsAg was 0.43% (23/5 398) vs 0.18%(8/4 410)in 2019, 0.66% (43/6 549) vs 0.20%(11/5 387)in 2020 and 0.41% (27/6 563) vs 0.09%(5/5, 615) in 2021. For initial screening, HBsAg deferral in 2021 was significantly different from 2019, while similar with 2020. 4) Among ALT deferral samples in the retest, 68.75% (11/16) were ALT≥45 U/L. Among HBsAg reactive samples, 91.67% (22/24) were reactive by single reagent. 【Conclusion】 Setting the threshold value of ALT for platelet donors in initial screening as less than 45 U/L can effectively reduce the reactive rate in the retest. HBsAg screening only for first-time platelet donors can reduce the detection cost. Adding pre-donation detection indicators according to local prevalence of transfusion transmitted diseases is conductive to reduce the discarding rate of platelets.

Characteristics analysis of HBV serological markers of NAT reactive blood donors under different HBsAg status / 中国输血杂志

【Objective】 To investigate the characteristics of HBV serological markers of NAT reactive blood donors under different HBsAg status. 【Methods】 NAT reactive samples, with HBsAg-, HBsAg+ /retest - and HBsAg+ by single reagent were collected from September 2021 to May 2022 in our laboratory. The TMA non-reactive samples were retested by Roche PCR, then HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc were detected by ECLI for statistical analysis. 【Results】 A total of 66 samples were collected, among which 55 were HBsAg-/NAT+. The positive rate of anti-HBc, anti-HBs+ anti-HBc, anti-HBe+ anti-HBc was 87.3% (48/55), 43.6% (24/55) and 45.5% (25/55), respectively. The positive rate of anti-HBs was 10.9% (6/55) and the overall negative rate was 1.8% (1/55). In 7 HBsAg+ initially/retest -/NAT+ samples, the positive rate of anti-HBc was 100%(7/7), and the positive rate of anti-HBe+ anti-HBc was 71.4%(5/7). In 4 HBsAg+ /NAT+ samples by single reagent, the positive rate of HBsAg+ anti-HBs+ anti-HBe+ anti-HBc was 50% (2/4), and positive rate of anti-HBe+ anti-HBc was100% (4/4). Samples, not reactive to TMA discriminatory and anti-HBc negative, were also non-reactive to individual PCR retest. There were significant differences in the positive rates of anti-HBe+ anti-HBc between HBsAg-/NAT+ samples and HBsAg+ /NAT+ (single reagent) samples (P<0.05). 【Conclusion】 Most HBsAg-/NAT+ blood donors were occult hepatitis B virus infection.The anti-HBe+ anti-HBc positive were correlated with HBV infection status. Non-reactivity discriminated by TMA plus anti-HBc negative do not exclude HBV DNA non-reactivity.

HBV infection among blood donors from 18 domestic blood stations of prefecture-level cities / 中国输血杂志

【Objective】 To analyze the hepatitis B virus (HBV) infection data of blood donors from 18 domestic blood stations, so as to investigate the HBV infection situation of blood donors. 【Methods】 The positive rate of HBV and its distribution characteristics of regions, the percentage of HBsAg+ ELISA in first-time vs repeated blood donors, and the percentage of HBsAg-/HBV DNA+ blood donors of 18 domestic blood stations during 2017 to 2020 were collected from the Working Platform for Practice Comparison of Blood Centers, and the HBV infection among blood donors were statistically analyzed. 【Results】 From 2017 to 2020, the positive rate of HBV in blood donors among 18 domestic blood stations was 13.48/10 000-144.02/10 000, with the average HBV positive rate in eastern, central and western region at 26.14/10 000, 51.98/10 000 and 41.00/10 000, respectively. The HBsAg+ rate by ELISA among first-time and repeated blood donors was 14.55/10 000-305.39/10 000 vs 1.04/10 000-87.43/10 000 The HBsAg-/HBV DNA+ yield was 1.80/10 000-35.31/10 000. 【Conclusion】 The distribution of HBV infection in blood donors has regional characteristics, and HBV prevalence was low in repeated blood donors. HBsAg ELISA combined with HBV DNA detection can better ensure blood safety.

Detection results of blood donors with HBsAg reactivity to single reagent: a retrospective analysis / 中国输血杂志

【Objective】 To retrospectively analyze the detection results of blood donors with HBsAg reactivity to single reagent detected by enzyme-linked immunosorbent assay (ELISA) in our center, so as to provide basis for further consolidating the blood donor team. 【Methods】 Samples of blood donors who had been deferred for at least 6 months due to HBsAg reactivity to sole ELISA assay were collected, and HBsAg ELISA and NAT were further performed. Meanwhile, HBsAg/HBsAb/HBeAg/HBeAb/HBcAb were detected by Roche electrochemiluminescence immunoassay, and the results were statistically analyzed. 【Results】 Among these 51 selected samples, 45 were negative to two assays, 6 were reactive to sole assay, with reactivity-yield rate at 11.76% (6/51). The results of NAT/ECLIA were all negative. For five indicators of hepatitis B virus infection, 23 samples were all negative and 28 were partially positive, mainly anti-HBs, anti-HBc and anti-HBe. 【Conclusion】 The follow-up detection of HBsAg ELISA sole-reagent reactive samples, supplemented with the detection of HBV serological markers, can reduce the number of deferred blood donors, increase the willingness to donate blood again, and protect the rights and interests of blood donors.

HBV laboratory tests and demographic characteristics in 169 897 first-time blood donors / 中国输血杂志

【Objective】 To investigate the HBV infection markers detection and demographic characteristics of first-time blood donors, so as to provide evidence for blood donor recruitment. 【Methods】 HBsAg was detected by ELISA, HBsAg negative samples were tested for HBV DNA, and chemiluminescence method was used to detect HBsAg, HBsAb, HBcAb, HBeAg and HBeAb in first-time blood donor HBsAg-/HBV DNA+ samples. Demographic information of HBsAg positive first-time donors was analyzed. 【Results】 From 2018 to 2022, a total of 502 739 people participated in voluntary blood donation, and first-time blood donors accounted for 33.79%. The HBsAg positive rate of first-time donors(28.37/10 000, 482/169 897)was higher than that of repeated blood donors(3.46/10 000, 115/332 842) (OR=8.23, 95%CI: 6.72~10.09), and the HBV DNA positive rate of first-time blood donors(4.83/10 000, 82/169 897)was lower than that of repeated blood donors(6.52/10 000, 217/332 842)(OR=0.74, 95%CI: 0.57~0.95). The positive rate of HBcAb in HBsAg-/HBV DNA+ samples of first-time blood donors was 73.17%. Significant differences were noticed in HBsAg-/HBV DNA+ and HBsAg positive rate among first-time blood donors among gender, age, education background and occupation (all P<0.05). 【Conclusion】 Low risk first-time blood donor recruitment is important for blood donation. Strengthening HBV screening before blood donation and detection after blood donation is beneficial to improve the safety of blood transfusion.

Prevalence of Hepatitis D Virus among People Attending the Antiretroviral Clinic and the Effect on the Liver Function Enzymes

Hepatitis D virus is an incomplete RNA virus requiring the assistance of the hepatitis B virus, specifically the HBsAg, to be infectious in humans. This study was designed to determine the prevalence of HDV among HIV patients and the effect on liver enzymes. The study was conducted at the Rivers state University Teaching hospital, Port Harcourt, Rivers State. Blood samples were obtained through vein puncture from 93 adults of which 41(44%) were males while 52(56%) were females between the ages 18 and 70 years attending the antiretroviral clinic and CD4+ cell count was also obtained. The samples were preserved at -20ºC. Each of the samples was tested using a SWE-Care rapid strip (China) for the presence of HBsAg. HDV antibody was detected using a Dia. Pro ELISA kit (Italy). The AST, ALT and ALP were determined. SPSS 21 was used to analyze the data and P values were determined. From the total samples collected, 7(7.5%) of them were positive to the HBsAg test of which 3(3.2%) were males, while 4(4.3%) of them were females. Of the 7 people positive to the HBsAg, 6(6.4%) were positive to the HDV antibody with 3(3.2%) females and 3(3.2%) males. There was significant depletion of the CD4+ cells across the groups. For the liver function test, the P values were ? 0.05 for AST, ALT and ? 0.05 for ALP. The HDV infection from the study was not gender, nor age based and suggests a negative impact on the CD4 cells. The liver function enzyme analysis, suggest higher risk of hypertension in HIV/HBV/HDV infection.

Demographic and Clinical Profile Analysis of Acute Viral Hepatitis

Background: Acute viral hepatitis (AVH) is a major health concern in developing countries like Bangladesh regarding morbidity as well as mortality. Usually, acute infections are caused by A, E, hepatitis viruses, and occasionally hepatitis B virus. Infection caused by the hepatitis C virus is usually asymptomatic. Prior knowledge of the demographic and clinical profile of acute viral hepatitis may be helpful for treatment professionals in the management of such patients. Aim of the study: The aim of this study was to assess the demographic and clinical profile of acute viral hepatitis patients in Bangladesh.Material & Methods:This prospective observational study was conducted in the Department of Gastroenterology, US-Bangla Medical College & Hospital during the period from March 2018 to February 2019, in Bangladesh. A total of 59 suspected patients with acute viral hepatitis were included as the study subjects for this study. Ethical approval of the study had been taken from the ethical committee of the mentioned hospital. A predesigned questionnaire was used in data collection. Collected data analyzed by using MS Office and SPSS version 23.0 programs as per need. A P-value, of <0.05 was considered significant.Results:Among 59 participants, the male-female ratio was 3.2:1 and the mean (±SD) age was 23.88±14.83 years. Analyzing hepatitis infection, we found hepatitis E was the highest in number 32(54.24%), followed by hepatitis E virus 20(33.9%), and hepatitis B virus 5(8.47%). In this study, we did not find any patients with hepatitis C virus, and 2 participants didn’t have any hepatotropic virus. The mean (±SD) Serum bilirubin (mg/dl) was 6.35±1.63, and the mean (±SD) serum alkaline phosphatase (IU/L), serum. creatinine (mg/dl), serum albumin (gm/dl) and plasma glucose random (mg/dl) were found 366.81±257.20, 6.97±14.96, 19.75±22.98 and 10.08±5.49 respectively. Considering dual viruses among the total of 5 patients along with hepatitis E & A viruses in this study, we found all were hepatitis B viruses. Among them, 3 with hepatitis E and the rest 2 were with hepatitis A virus consequently. Among them 3(60%) with hepatitis A and the rest 2(40%) with hepatitis E. In the issue of patients attended with clinical complaints, vomiting was the highest among hepatitis A patients 19(86.4%) followed by jaundice 8(36.4%) and fever8(36.4%). On the other hand, among hepatitis E patients’ jaundice was highest at 19(61.3%) followed by vomiting at 17(54.8%), and fever at 6(19.4%) respectively.Conclusions:The incidence of HEV is found as the most predominant among all the acute viral hepatitis patients and vomiting and jaundice were the most common presenting complaints.

Hepatitis D virus seroprevalence in HBsAg positive patients attending a tertiary care hospital in Northeast India

Background:Hepatitis D virus (HDV) infection is present worldwide. Around 18 million people are estimated to be infected with HDV and can infect individuals with active HBV infection and cause severe liver disease. There is lack of data on the prevalence of HDV infection in the state and also in the region. The aim of the study was to determine the seroprevalence of HDV in HBsAg positive patients attending Regional Institute of Medical Sciences Hospital, Imphal, Manipur, India.Methods:This study was carried out in a tertiary care hospital (Regional Institute of Medical Sciences, Imphal).The study was done for a period of 2 years from September 2016 to August 2018. A total of 119 HBsAg ELISA positive cases were included in the study.Results:Out of 119 HBsAg positive cases, 5 cases were positive for hepatitis D antibodies, of which 3 were positive for anti-Hepatitis D virus IgM and 2 were positive for anti-Hepatitis D virus IgG.Seroprevalence of HDV infection was found to be 4.2%.Conclusions:Seroprevalence of HDV infection was found to be 4.2% which is higher than the finding in some of the recent studies in the country.

Hepatitis B: Anticuerpos contra antígeno de superficie en residentes de pediatría, Hospital Central de Maracay

Introducción: Los profesionales del área de la salud tienen un riesgo incrementado de contraer la infección por el virus de hepatitis B (VHB). Objetivo: Evaluar anticuerpos contra el antígeno de superficie de la hepatitis B, en los residentes de pediatría del Hospital Central de Maracay en el período junio-agosto de 2021. Materiales y métodos: Estudio clínico epidemiológico, no experimental y de corte transversal, en el que se tomó muestra sanguínea a 54 médicos residentes para la determinación de anticuerpos contra el antígeno de superficie del VHB (Anti-HBs). Resultados: El promedio de edad fue 27,48 años con una desviación estándar de 1,6. El 83,33 % pertenecían al sexo femenino, 51,85.% cursaban el 1er año del posgrado, 33,33 % con esquema de vacunación documentado, de estos, 66,67.% completaron el esquema y 77,78 % cumplidos en la adultez. Con respecto al tiempo de la última dosis, el 66,67 % hasta 10 años. Se detectaron niveles de Anti-HBs mayores de 10 mUl/mL en el 94,44 %, con mayor prevalencia de niveles protectores a favor del sexo femenino. Se evidenció una correlación lineal positiva entre los niveles de Anti-HBs y el tiempo desde la última dosis de la vacuna contra la hepatitis B. Conclusiones: Aunque existe una debilidad en los médicos residentes en cuanto a la tenencia y cumplimiento del esquema de inmunización, la mayoría de ellos mostraron niveles protectores de anti-HBs. A mayor tiempo transcurrido desde la última dosis de la vacuna hay un descenso en los niveles de anti-HBs lo que justifica dosis de refuerzo a los 10 años.

Introduction: Health professionals have an increased risk of contracting hepatitis B virus infection (HBV). Objective: To evaluate antibodies against hepatitis B surface antigen in residents of pediatrics of the Central Hospital of Maracay in the period June-August. 2021. Materials and methods: Clinical epidemiological, nonexperimental and cross-sectional study, in which blood samples were taken from fifty-four medical residents for the determination of antibodies against the HBV surface antigen. Results: The average age was 27.48 years with a standard deviation of 1.6. 83.33 % were female, 51.85 % were in the first year of postgraduate studies, 33.33 % had a documented vaccination schedule, of these, 66.67 % completed the schedule and 77.78 % completed it in adulthood. Regarding the time of the last dose, for 66.67 % of the study population, it was up to 10 years ago. Anti-HBs levels greater than 10mUl/ml were detected in 94.44 %, with a higher prevalence of protective levels in favor of the female sex. A positive linear correlation between the levels of Anti-HBs and the time since the last dose of the hepatitis B vaccine was evidenced. Conclusions: Although there is a weakness in the resident doctors in terms of possession and compliance with the immunization schedule, the most of them showed protective levels of anti-HBs. The longer the time elapsed since the last dose of the vaccine, there is a decrease in anti-HBs levels, which justifies a booster dose at 10 years.

Prevalence of incidental and total human immunodeficiency virus, hepatitis B and hepatitis C seropositivity among patients posted for cataract surgery at a tertiary care center in India

Purpose: To study the prevalence of human immunodeficiency virus (HIV), hepatitis B (HBV), and hepatitis C (HCV) viral seropositive among the patients posted for cataract surgery at a tertiary care center in north India. Methods: It was a cross?sectional study done for 30 months duration. All the patients posted for cataract surgery underwent comprehensive ophthalmic evaluation followed by routine hematological workup, including viral markers for HIV, Hepatitis B surface antigen (HBsAg), and anti?HCV. Data were analyzed by the Statistical Package for Social Science (SPSS Version 20). Results: A total of 7,316 individuals underwent cataract surgery from Jan 2016 to August 2018, 4,073/7,316 (55.7%) were males. The prevalence for HIV was 58/7,316 (0.8%), HBsAg was 151/7,316 (2.1%), and HCV was 11/7,316 (0.1%); 28/58 (48.3%) HIV positives were unaware of their seropositivity till testing, as were 37/151 (24.5%) of HBsAg positives, and 4/11 (36.4%) HCV positives. There was a significant relationship between the mean age in the patients with HIV (P = 0.002) and anti?HCV (P = 0.045). A majority of the seropositive patients were found to be illiterate (45.6%), followed by educated up to high school level (29.1%), and graduate (25.0%). Conclusion: Viral seropositivity was significant among the patients posted for cataract surgery. The eye care providers could refer these patients for counseling and further management for the patient’s and their caretaker’s benefit

A cross-sectional survey on the epidemiology of HBV infection among HBsAg-positive people in Deyang Area / 公共卫生与预防医学

Objective To analyze and discuss the epidemiological characteristics of HBV infection in people with hepatitis B virus (HBV) surface antigen (HBsAg) positive in Deyang Area. Methods The health checkup information of the region in 2020 was selected to screen the subjects with the results of HBV serological markers, and 1645 HBsAg-positive patients were selected as the main subjects to analyze the epidemiological characteristics of the population. Use Epidata to enter data, and use SPSS statistical software package to analyze the data. Results Except for the differences in occupational distribution between the sexes, the differences in age, education level, family history, and vaccination history (hepatitis B vaccine) were not statistically significant among men and women in this study. Among the 1645 HBsAg-positive subjects, 41 were double-positive for anti-HBs at the same time, and the double-positive rate was 2.49%. Compared with HBs-negative people, HBs-positive people have a relatively higher proportion of males, junior high school education level, occupation as a farmer, unvaccinated, and positive family history. AmpliPrep/COBAS quantitative detection reagent was used for HBV DNA viral load detection, 92.69% were quantitatively positive (38 cases), of which 23 cases (60.53%) with HBV DNA 20 IU/mL). The epidemiological characteristics of the double-positive population suggest that the proportion of males, junior high school education level, occupation as a farmer, unvaccinated and positive family history is relatively higher. Analyzing the potential influencing factors of anti-HBs positive (double positive) in HBsAg-positive people, it can be seen that the increase in age, the occupation is a farmer, the first injection of hepatitis B vaccine is not in time, and the family history of liver disease is positive as potential risk factors for the occurrence of double positive. Conclusion In the future, this region should pay special attention to farmers who are older, have a family history of liver disease, and lack or incomplete hepatitis B vaccination for hepatitis B prevention and control.

Are there only a few steps away from the prime time for hepatitis B virus RNA as a routine marker to guide decision making in treatment of chronic hepatitis B? / 临床肝胆病杂志

This paper discusses HBsAg and HBV RNA as routine markers to guide treatment decisions of chronic hepatitis B.

Cytokine Profiles and Virological Marker Monitoring during 48 Weeks Peginterferon Alfa Treatment for HBeAg-Positive Chronic Hepatitis B / 生物医学与环境科学（英文）

Objective@#This study aimed to investigate whether cytokine profiles and virological markers might add value in monitoring the effects of peginterferon (PEG-IFN) therapy for hepatitis B e-antigen (HBeAg) positive chronic hepatitis B (CHB).@*Methods@#HBeAg positive patients with CHB were treated with PEG-IFN for 48 weeks. Clinical biochemical, and HBV serological indexes, as well as cytokines, were detected at baseline and every 12 weeks.@*Results@#A total of 116 patients with CHB were enrolled in this study; 100 patients completed the 48-week treatment and follow-up, of whom 38 achieved serum HBeAg disappearance, 25 achieved HBeAg seroconversion, 37 showed HBsAg decreases ≥ 1 log 10 IU/mL, 9 showed HBsAg disappearance, and 8 became HBsAb positive. The cytokine levels at baseline and during treatment were similar between the HBeAg disappearance group and non-disappearance group. The disappearance of HBeAg was independently associated with HBeAg levels at weeks 12 and 24, and with the HBeAg decline at week 24 ( P < 0.05). The HBsAg response was independently associated with HBsAg, the HBsAg decline, HBeAg, the HBeAg decline at week 12, and HBsAg at week 24 ( P< 0.05).@*Conclusion@#There was no significant correlation between the response to interferon (IFN) and cytokines during PEG-IFN treatment. The changes in virological markers predicted the response to IFN after 48 weeks.

Serological and molecular characterization of HBV infection with HBsAg reactive and HBV DNA non-reactive / 中国输血杂志

【Objective】 To investigate the serological and molecular characteristics of HBsAg+ /HBV DNA non-reactive (NR) infections. 【Methods】 Samples tested as HBsAg+ and HBV DNA NR were confirmed by individual NAT repeat testing, viral particle concentration by PEG precipitation combined with in-house nested PCR and real-time quantitative PCR, anti-HBc testing, and HBsAg quantification. HBV sequences were compared with those from donors with chronic and occult infection as controls. 【Results】 A total of 792 195 samples were screened between January 2011 and December 2020, of which 53 (1: 14 947) were confirmed HBsAg+ /HBV DNA NR. HBV DNA was detected further in five (9.4%) samples; three S sequences and four Pre Core/Core sequences were obtained. Unique amino acid substitutions (P130T, P135Q/S, R151Q, G153S and S155F) were found in the Core protein that may affect virus packaging and replication. 【Conclusion】 Extremely low HBV DNA level was detected in plasmas of HBsAg+ /HBV DNA NR donors. Barely detectable HBV DNA might be associated with unusual mutations in the Pre Core/Core protein affecting viral replication. More sensitive HBV DNA and/or HBsAg assays may be considered to further reduce the potential HBV transfusion-transmission residual risk.

Evaluation of CLIA plus NAT for detecting HBsAg single-ELISA-reactive samples of blood donors / 中国输血杂志

【Objective】 To study the detection performance of HBsAg single-ELISA-reactive samples of blood donors. 【Methods】 Two kinds of ELISA reagents from different manufacturers (named as reagents A and B) were used for HBsAg screening. A total of 276 samples, from January 2017 to May 2021, with HBsAg single-ELISA-reactive results were collected for further nucleic acid detection technology (NAT) and chemiluminescence immunoassay (CLIA) testing, to undergo HBV-DNA and five hepatitis B tests, respectively. The relationship between HBsAg single-ELISA-reactivity, NAT and CLIA was statistically analyzed. 【Results】 Among the 276 HBsAg single-ELISA-reactive samples, 14 were NAT reactive, with the positive rate of 5.07% (14/276). Fisher′s exact test was used to compare the compliance of reagents A and B with NAT reactivity, and the difference was not statistically significant (P<0.05). Among 14 HBsAg+ /NAT+ samples retested by CLIA, 2 were HBsAg reactive(14.29%, 2/14), 13 were anti-HBc reactive (92.86%, 13/14), 9 had the quantitative value of anti-HBs <10 mIU/mL, 5 had the quantitative value of anti-HBs between 10 to 100mIU/ mL. A total of 5 serological patterns were detected, and anti-HBe+ /anti-HBc+ pattern was the dominant. There were 262 cases of HBsAg+ /NAT- samples, but only 1 (0.38%, , 1/262) case was HBsAg reactive by CLIA, 100 were anti-HBc reactive (38.17%, 100/262), 144 (54.96%, 144/262) were anti-HBs reactive, and 1 was HBeAg reactive. A total of 8 serological patterns were detected. 【Conclusion】 Most of HBsAg single-ELISA-reactive results are false, and NAT could effectively reduce the residual risk of transfusion transmitted diseases.

Application of serum plate in the grey area setting of HBsAg ELISA / 中国输血杂志

【Objective】 To evaluate the necessity and rationality of setting 0.7 CO of the gray area of HBV surface antigen reagent (ELISA) in our laboratory, and to provide the basis for the grey area setting. 【Methods】 A total of 60 samples of serum plate were repeatability tested by two kinds of ELISA HBsAg reagents. Based on the test results, C50, C5, C95 concentrations and corresponding S / CO values were calculated, and whether C5 and C95 were within C50±20% was verified. At the same time, the true positive rate and the confirmed positive rate of gray area samples revealed by two reagents were calculated. 【Results】 The confirmation results of serum plate were as follows: The C50, C5 and C95 concentrations of reagent A were 0.090 IU/mL, 0.075 IU/mL, 0.105 IU/mL. The true positive rate was 99.1%(436/440), the confirmed positive rate of grey area was37.8%(136/360). The C50, C5 and C95 concentrations of reagent B were 0.112 IU/mL, 0.091 IU/mL, and 0.133 IU/mL; the true positive detection rate was 97.1%(233/240); the confirmed positive rate of grey area was 35.8%(129/360). It was verified that C5 and C95 of reagent A and reagent B were within their respective C50±20%, and the gray areas of the HBsAg ELISA were verified to be effective. 【Conclusion】 It is necessary to set the gray area for these two HBsAg ELISA reagents in our laboratory, but the gray area value setting to 0.7 CO is unreasonable. The best gray area value was 0.75 CO in reagent A and 0.63 CO in reagent B.

Significación de los marcadores infecciosos para identificar portadores de hepatitis B en donantes de sangre / Significance of hepatitis B surface antigen, IgM/IgG core antibody and hepatitis B virus DNA in blood donors

Resumen Introducción: La identificación de portadores del virus de la hepatitis B en donantes de sangre es imperativo para evitar la transmisión de la enfermedad a través de transfusiones sanguíneas. Objetivo: Determinar si los donantes de sangre con resultados positivos de los marcadores serológicos HbsAg y anti-HBc eran portadores de ADN del virus de la hepatitis B. Métodos: Se recolectaron 12 745 muestras de seis bancos de sangre ecuatorianos, las cuales fueron analizadas con pruebas serológicas para identificar los marcadores infecciosos HBsAg, anti-HBc, anti-HBs mediante prueba ELISA automatizada. Todas las muestras positivas para uno, dos o los tres marcadores fueron analizadas con técnica molecular para determinar la presencia de ADN viral. Resultados: Se identificó que 27.5 % de las muestras reactivas solo a anti-HBc y 100 % de las muestras con resultados positivos de HBsAg/anti-HBc-IgM/IgG presentaron ADN del virus de la hepatitis B (p = 0.001). Conclusiones: La elección de los marcadores de infección y los métodos de detección definen los resultados. Es importante la realización de dos pruebas serológicas y una molecular para identificar a los portadores del virus de la hepatitis B y evitar su transmisión.

Abstract Introduction: Identification of hepatitis B virus carriers in blood donors is imperative in order to avoid transmission of the disease via blood transfusion. Objective: To determine if blood donors with positive results for serological markers HBsAg and anti-HBc were hepatitis B virus DNA carriers. Methods: 12,745 samples were collected from six Ecuadorian blood banks and analyzed for HBsAg, anti-HBc and anti-HBs infectious markers by automated ELISA. All samples that tested positive for one, two or all three markers were analyzed with molecular techniques to determine the presence of viral DNA. Results: 27.5 % of the samples that were reactive for anti-HBc alone and 100 % of those with positive results for HbsAg and IgM/IgG anti-HBc were identified to contain hepatitis B virus DNA (p = 0.001). Conclusions: The selection of infection markers, as well as the detection methods define the results. Performing two serological and one molecular test is important in order to identify hepatitis B virus carriers and prevent its transmission.

Risk factors for losing hepatitis B virus surface antibody in patients with HBV surface antigen negative/surface antibody positive serostatus receiving biologic disease-modifying anti-rheumatic drugs: a nested case-control study

Abstract Background: Hepatitis B virus (HBV) reactivation consequent to immunosuppressive therapy is an increasingly prevalent problem with serious clinical implications. Treatment with biologic agents conduces to the loss of protective antibody to HBV surface antigen (anti-HBs), which significantly increases the risk of HBV reactivation. Hence, we investigated the risk factors for losing anti-HBs in patients with rheumatic diseases and HBV surface antigen negative/anti-HBs positive (HBsAg-/anti-HBs+) serostatus during treatment with biologic disease-modifying anti-rheumatic drugs (DMARDs). Methods: Using a nested case-control design, we prospectively enrolled patients with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis/psoriasis, or juvenile idiopathic arthritis, who were treated with biologic DMARDs at Changhua Christian Hospital, Taiwan, from January 2013 to June 2019 and had HBsAg-/anti-HBs+ serostatus; the analytic sample excluded all patients with HBsAg+ or anti-HBs- serostatus. Anti-HBs titers were monitored 6-monthly and cases were defined as anti-HBs < 10 mIU/ml during follow-up. Cases were matched one- to-all with controls with anti-HBs ≥ 10 mIU/ml on the same ascertainment date and equivalent durations of biologic DMARDs treatment (control patients could be resampled and could also become cases during follow-up). Between-group characteristics were compared and risk factors for anti-HBs loss were investigated by conditional logistic regression analyses. Results: Among 294 eligible patients, 23 cases were matched with 311 controls. The incidence of anti-HBs loss was ∼ 2.7%/person-year during biologic DMARDs treatment. Besides lower baseline anti-HBs titer (risk ratio 0.93, 95% CI 0.89-0.97), cases were significantly more likely than controls to have diabetes mellitus (risk ratio 4.76, 95% CI 1.48-15.30) and chronic kidney disease (risk ratio 14.00, 95% CI 2.22-88.23) in univariate analysis. Risk factors remaining significantly associated with anti-HBs loss in multivariate analysis were lower baseline anti-HBs titer (adjusted risk ratio 0.93, 95% CI 0.88-0.97) and chronic kidney disease (adjusted risk ratio 45.68, 95% CI 2.39-871.5). Conclusions: Besides lower baseline anti-HBs titer, chronic kidney disease also strongly predicts future anti-HBs negativity in patients with HBsAg-/anti-HBs+ serostatus who receive biologic DMARDs to treat rheumatic diseases. Patients with low anti-HBs titer (≤ 100 mIU/ml) and/or chronic kidney disease should be monitored during biologic DMARDs therapy, to enable timely prophylaxis to preempt potential HBV reactivation.