Detection Of HBV Research Articles

Simple and Feasible Detection of Hepatitis B Virus via Combination of Multienzyme Isothermal Rapid Amplification and Lateral Flow Dipstick Strip.

Hepatitis B virus infection is not only a huge burden in the field of social health but also a major public health problem that affects the lives and health of the people. Simple, rapid, feasible detection of HBV is critical for its prevention and spread, especially in the developing countries with low-resource laboratories. To this end, we combined multienzyme isothermal rapid amplification (MIRA) and lateral flow dipstick (LFD) strip to detect HBV. A pair of primers targeting the conserved region of HBV genome was designed and used in MIRA-LFD assay. Our results found that the entire amplification of MIRA-LFD only takes 10 min at 37°C and the dilution of the amplification products was added in the LFD strip and observed by the naked eye after 10 min. The detection sensitivity of this method can reach 10 pg. The 45 clinical samples were detected by MIRA-LFD and real-time PCR. The accuracy rate of MIRA-LFD was 100%. Therefore, these characteristics of our newly developed MIRA-LFD assay make it particularly useful and suitable for detecting HBV in the resource-limited condition.

Comparative evaluation of ADVIA Centaur® XP chemiluminescence system for screening of HBV, HCV, HIV and syphilis in Indian blood donors

BackgroundThe introduction of chemiluminescent immunoassay (CLIA) in blood donor screening has led to a gradual replacement of enzyme-linked-immunosorbent assay (ELISA) as the former offers automation, higher sensitivity and lower turn-around-time. However, only a few CLIA platforms are used for blood donor screening in India. The present study evaluated one such newer platform viz., Adiva Centaur XP CLIA for screening of HBV, HCV, HIV and syphilis. Material and methodsProspective comparative study wherein 4843 whole blood donors were screened for HBsAg, Anti-HCV, HIV Ag-Ab and Anti-treponemal antibodies in both Advia Centaur XP and Architect i2000SR platforms. Additional tests were performed in samples which were reactive in only one of the platforms. The sensitivity, specificity, positive predictive value, negative predictive value, accuracy, false positive rate and false negative rate of both the platforms were compared. Kappa coefficient was calculated to determine the agreement between the testing platforms. ResultsThe sensitivity of Advia Centaur platform for HBV, HCV, HIV and syphilis detection were 94.9 %, 100 %, 100 % and 100 % respectively as compared to 96.6 %, 100 %, 100 % and 100 % in Architect i2000SR platform. The specificity of both the platforms were 99.8−99.9 % for all the four tests. The agreement between the two platforms was almost perfect for HBV, HCV and syphilis testing; and fair for HIV testing. ConclusionThe Advia Centaur CLIA platform was found to be comparable with the Architect CLIA platform for blood donor screening. Unexpected finding was the occurrence of HBV false negatives in both the platforms, possibly due to HBsAg mutations.

Amplification-free smartphone-based attomolar HBV detection

Classical gold standard HBV detection relies on expensive devices and complicated procedures, thus is always restricted in large-scale hospitals and centers for disease control and prevention. To extend HBV detection to primary clinics especially in underdeveloped areas, we design amplification-free smartphone-based attomolar HBV detecting technique based on single molecule sensing. Verified by synthesized HBV target DNA, this technique reaches a detection limit at attomolar concentration (100 aM); and verified by 110 clinical samples, it also reaches a rather high sensitivity of 104 copy/mL (≈2000 IU/mL) with a high accuracy of 93.64% certificated by gold standard HBV detecting devices. Besides, this technique can quantify HBV viral load in 70 min only using portable and inexpensive devices as well as simple operations. Because of its cost-effective, field-portable and operable design, highly sensitive and selective detecting capability and wireless data connectivity, this technique can be potentially used in mobile HBV diagnoses and share HBV epidemic information especially in resource limited situations.

Serodiagnosis of Hepatitis B Virus Using PreS Antigen from Pakistani Isolate SBS001

Background: An estimated 325 million people worldwide live with hepatitis B and/or C and approximately, 5 million people are affected with hepatitis B. in Pakistan.This study aimed at developing PreS protein from Hepatitis B Virus Pakistani isolate (SBS001) with enhanced sensitivity to detect antibodies in serum as a diagnostic method.
 Methods: Gene encoding PreS region from hepatitis B Virus was cloned and expressed in Escherchia coli. The recombinant protein preS-His was purified by Ni-IDA affinity chromatography. Antibodies were raised in rabbit. This protein was screened for detection of antibodies in HBV patients’ sera through ELISA. This ELISA procedure was compared with commercially available Kit used for diagnosis of HBV infection.
 Results: Single band purified recombinant PreS protein was obtained with high titer of antibodies raised in rabbits. This recombinant protein was used in ELISA as antigen coated on the plate. That efficiently detected antibodies present in HBV patients. It was concluded that preS-His antigen/ protein A HRP-conjugate ELISA method was more sensitive than the commercial kit for detecting the antibodies present in HBV patient sera.
 Conclusion: It was concluded that SBS001 PreS recombinant protein can be used in ELISA kits for detection of HBV in Pakistani population.

Clinical and Histological Long-Term Follow-Up of De Novo HBV-Infection after Liver Transplantation.

Background and Objectives: Development of hepatitis-B is considered a serious complication after liver transplantation. HBV de novo infection is a rather rare phenomenon, however it deserves attention in the era of donor organ shortage. The aim of the present analysis was to examine its course in liver transplant patients. Materials and Methods: Prevalence of de novo HBV-infections was extracted from our local transplant data base. Analysis focused on the moment of HBV-detection and on the long-term follow-up in terms of biochemical and histological changes over 30 years. Results: 46 patients were identified with the diagnosis of de novo hepatitis B. Median time from liver transplantation to diagnosis was 397 days (7–5505). 39 patients received antiviral therapy. No fibrosis progression could be detected, whereas the grade of inflammation significantly lessened from the moment of HBV detection to the end of histological follow-up over a median of 4344 days (range 123–9490). Patients with a poor virological control demonstrated a significantly poorer overall survival. Conclusions: De novo hepatitis B in liver transplant patients is a condition that can be controlled very well without significant fibrosis progression or graft loss if recognized on time within a regular transplant follow-up schedule.

CRISPR/Cas12-Based Ultra-Sensitive and Specific Point-of-Care Detection of HBV.

Hepatitis B remains a major global public health challenge, with particularly high prevalence in medically disadvantaged western Pacific and African regions. Although clinically available technologies for the qPCR detection of HBV are well established, research on point-of-care testing has not progressed substantially. The development of a rapid, accurate point-of-care test is essential for the prevention and control of hepatitis B in medically disadvantaged rural areas. The development of the CRISPR/Cas system in nucleic acid detection has allowed for pathogen point-of-care detection. Here, we developed a rapid and accurate point-of-care assay for HBV based on LAMP-Cas12a. It innovatively solves the problem of point-of-care testing in 10 min, particularly the problem of sample nucleic acid extraction. Based on LAMP-Cas12a, visualization of the assay results is presented by both a fluorescent readout and by lateral flow test strips. The lateral flow test strip technology can achieve results visible to the naked eye, while fluorescence readout can achieve real-time high-sensitivity detection. The fluorescent readout-based Cas12a assay can achieve HBV detection with a limit of detection of 1 copy/μL within 13 min, while the lateral flow test strip technique only takes 20 min. In the evaluation of 73 clinical samples, the sensitivity and specificity of both the fluorescence readout and lateral flow test strip method were 100%, and the results of the assay were fully comparable to qPCR. The LAMP-Cas12a-based HBV assay relies on minimal equipment to provide rapid, accurate test results and low costs, providing significant practical value for point-of-care HBV detection.

Screening of Occult Hepatitis B Virus Infection among Egyptian Blood Donors.

Background:Although sensitive screening assays for hepatitis B virus surface antigen (HBsAg) are available, special cases of post-transfusion hepatitis B virus infection still occur. The present study was conducted to evaluate the prevalence of anti-hepatitis B core (anti-HBc) positivity and the presence of HBV-DNA in serum samples of healthy blood donors negative for both HBsAg and anti-HCV antibodies in Benha, Egypt. Materials and methods: The study included a screening of 450 selected blood donors. The distribution of blood donors was 288 males (64%) and 162 females (36%). The recruited blood donors who met the criteria for blood donation were routinely screened for HBsAg, HIV I/2-Ab, and syphilis-antibodies. The blood units for donations were further analyzed for the presence of HBc-IgM and HBV-DNA levels by PCR method. Results: Testing of the accepted units for the donation was about 12 (2.7%) HBc-IgM positive, and 9 (2%) HBV-DNA positive units. The standard screening of blood unit failed to recognize early acute or window HBV infection where HBsAg is missing. Conclusions:Our investigations proposed that sensitive methods for the detection of HBV by PCR might be recommended in the screening of donated blood. Furthermore, anti-HBc antibodies should be tested regularly on all blood donation units.

Method for detecting hepatitis B virus in blood plasma at low viral load using real-time PCR.

A method for detecting HBV DNA in peripheral blood at low viral load using real-time PCR was developed and its significance in identifying HBsAg-negative viral hepatitis B was evaluated. When developing the method, blood plasma samples and liver tissue biopsy material were used from 128 patients living in St. Petersburg, in various regions of the Russian Federation, as well as in the Central Asia countries. We also used blood plasma samples from 96 pregnant women and 37 hemodialysis center patients living in Northwestern Federal District, 199 foreign citizens undergoing medical examination to obtain work permits at the Directorate for Migration in the Northwestern Federal District, 397 conditionally healthy people living in the Socialist Republic of Vietnam. HBV was detected by nested PCR. Analytical sensitivity was tested using the stepwise dilution method. According to the method developed by us, at the first stage, the HBV DNA is amplified using at the first stage oligonucleotides flanking the genome region 2932-3182 ... 1-1846 nt., and at the second stage two oligonucleotides pairs to the genome virus regions (gene S and gene X) and corresponding oligonucleotide fluorescently labeled probes complementary to the amplified fragments regions carrying fluorophores at the 5'-end, and non-fluorescent quenchers at the 3'-end. The channel corresponding to the FAM fluorophore detects the HBV DNA S-region amplification product, and the channel corresponding to the ROX fluorophore detects the HBV DNA X-region amplification product. The method sensitivity for DNA extraction from plasma with a 100 μl volume was 10 IU/ml. Obtaining a threshold cycle Ct for only one FAM or ROX fluorophore may indicate the HBV DNA presence in a sample at a load of less than 10 IU / ml, HBV detection in this case is possible with a repeated PCR study of the corresponding sample with HBV DNA extraction from an increased plasma volume (200-1000 μl). The developed method makes it possible to identify various HBV genovariants, both characteristic and rare in the Russian Federation, circulating in other world regions. The method can be used to detect HBV in risk groups, in the population, as well as in screening blood donors in order to ensure the blood transfusions safety.

Comparison of ELISA and NAT techniques among blood donors

Large number of blood transfusions are carried out globally to save countless lives, but unsafe practices risks the recipient of transfusion transmittable infections ((TTI’S). TTI’s can be reduced by improving donor selection, donor awareness regarding TTI and implementing sensitive screening tests. In India, it is mandatory to test each donated unit of blood for markers of HIVI and II, HBV, HCV, malaria and syphilis. Currently, in India all the blood donations are screened for various infectious markers using ELISA or rapid methods. TTIs can be responsible for causing considerable burden on the health status and economy of the country. Hence there is need to introduce better methods for testing blood units. Even after being seronegative the blood transfusions are still at risk of transmitting infections. In order to reduce the residual risk sensitive screening tests are needed and as a result NAT has been considered. In India NAT testing is not mandatory but many private blood banks and hospitals and state Governments of few states have started implementing NAT for blood safety. It increases the possibility of identifying the infection in window period and thus reducing the residual risk of TTIs. NAT is highly sensitive and specific for viral nucleic acids and is based on amplification of targeted regions of RNA and DNA and thus is the technique of choice. By early detection than serology, the window period of HIV, HBV and HCV infections narrows. But the issue of higher cost accounts for its limited use especially in developing countries. The present study is designed to understand the role of NAT in detecting the risk acquiring TTIs as compared to conventional methods currently in use for detection of HBV, HCV and HIV infections amongst blood donors.

Serological and Molecular Detection of Hepatitis B virus among patients referred to Kurdistan Center for Hepatology and Gastroenterology in Sulaimani City/Kurdistan Region of Iraq

Hepatitis B virus infection is caused by the hepatitis B virus, a major global health problem. This infection can lead to chronic conditions, followed by cirrhosis and hepatocellular carcinoma (HCC). The current study was aimed to detect HBV using serological and molecular techniques. During 2019, 300 blood samples were collected from Kurdistan Center for Hepatology and Gastroenterology in Sulaimani city. Enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) techniques were used for the detection of HBsAg and HBV DNA, respectively. Obtained results were revealed that 92 out of 300 tested patients (30.66%) seropositive for HBsAg. Among 92 seropositive patients, 53 were shown positive results for HBV DNA by RT-PCR. Dental clinic visiting and dialysis were among the important risk factors for HBV transmission. The vast majority of positive results were among males. Smokers showed relatively high rates of positive results. One-third of the referred patients who had liver complaints were positive for HBsAg. More than half of the seropositive patients showed RT-PCR positive results. It was concluded that the molecular method (RT-PCR) is more sensitive and gives a more accurate result than serology (ELISA). Therefore, it can be used as a diagnostic tool for HBV detection.

Hepatitis B virus infection among Nigerian pediatric population

ABSTRACT Hepatitis B infection has continued to be a growing global health concern in spite of the discovery and administration of hepatitis B vaccines over the years. The focus of this study was to evaluate the burden of this infection among asymptomatic pediatric population. Children, who visited the hospital between March and August 2018, were recruited into this study. Blood samples were collected after obtaining informed consent. The samples were processed and screened for HBsAg. Data were analyzed using chi-square statistic. Six (2.9%) of the 206 children in the study were positive for HBsAg. A hundred (48.5%) of the children were males and 106 (51.5%) were females in the ratio 1:1.1 (male to female). More male tested positive (4%) (4/96) than their female counterparts (1.9%) (2/104)(x2 = 0.813; p = .367). Age group 13–18 years had the highest infection rate 6.6% (4/57), followed by age group 5–12 years with 3.8% (2/50). No HBV detection was recorded among those below the age of 5 years. Patients with no vaccination history had a higher HBV infection rate (4.2%) compared with those who had been vaccinated (1.1%) (x2 = 1.164; p = .281). The study showed that HBV infection is of intermediate endemicity among the study population, a reflection of the endemic status of the infection in Nigeria. Vaccination effort among children should be sustained and intensified. We advocate for a more intensive public enlightenment on the infection especially among women of marriageable age.

A mixed-methods approach to understanding perceptions of hepatitis B and hepatocellular carcinoma among ethnically diverse Black communities in South Florida.

Hepatitis B (HBV), the leading cause of hepatocellular carcinoma (HCC), disproportionately affects minorities. Compared to other races, Blacks more often present with advanced HCC and have decreased survival. We observed higher HBV-associated HCC rates among Blacks than reported nationally. In our center, Haitian Blacks had the highest rates of HBV-associated HCC and shorter survival compared to other Blacks. We investigated knowledge and perceptions regarding HBV and HCC among Blacks born in the United States or Haiti. Using community partnerships, participants were recruited via word of mouth, email, social media or from Hepatology clinic. Focus groups were conducted in Haitian Creole or English and stratified by birthplace, gender and infection status. Discussions were audio-recorded and transcribed verbatim. A constant comparative method was used for data analysis; themes are based on conversational details. There were 55 participants; 49% were male and 27% had chronic HBV. Only 42% of Haitian Blacks knew about HBV prior to participation vs. 78% of African Americans, p 0.03. Both groups expressed that fear, mistrust of the medical establishment, denial and stigma might compel persons to avoid seeking care. Both groups attributed higher rates of late stage HCC diagnosis in Blacks to inadequate financial resources and education. Those with HBV reported confusion regarding their infection and suboptimal communication with healthcare providers. In two communities disproportionately affected by HBV, misconceptions about disease transmission, stigma, low health literacy and decreased access to care may limit detection for HBV. Culturally relevant community-based interventions are needed to increase HBV detection.

Hepatitis B Virus Nucleic Acid Amplification Testing Assay in Detecting Window Period and Occult Hepatitis B Virus Infections in Blood Donors

SummaryTo reduce the residual risk of transfusion-transmitted infections, nucleic acid amplification testing (NAT) of donated blood with higher sensitivity for HBV, HCV, and HIV 1/2 was implemented in Bulgaria at the end of 2019. This study aimed to assess the clinical sensitivity of HBsAg testing and NAT testing of donated blood to detect all forms of HBV infection. A total of 9498 consecutive blood donations collected for six months, from February 10 to July 17, 2020, from first-time and repeat donors at the Regional Center of Transfusion Hematology Pleven, Bulgaria, were screened for HBsAg and HBV DNA. The detection of HBsAg was performed by enzyme-linked immunoassay and chemiluminescent immunoassay. Detection of HBV DNA was performed using the HIV1/2 /HCV / HBV multiplex Procleix Ultrio Elite assay in a fully automated and integrated Procleix Panther System. The overall HBsAg prevalence was 0.05%. HBV DNA was detected in 25 blood units (0.26%), but only 12 (0.13%) were found positive after repeat testing and were confirmed by a discriminatory test. The other 13 units were false positive, with the initial reactive result and negative results after repeat testing. HBV DNA’s overall incidence was significantly higher in HBsAg-positive donors than the HBsAg- negative (Fisher exact p=0.0063). In our study, blood donations were not tested for anti-HBc and anti-HBs, so it is difficult to determine whether HBV DNA-positive/HBsAg-negative results were associated with the early phase of infection or persistent occult infection. There was no statistical difference in the incidence of HBV DNA between repeat-donors (0.16%) and first-time donors (0.06%) (Fisher exact test p=0.239 NS), and also between the incidence in female donors (0.12%) and male donors (0.13%) (Fisher exact test p=1.0 NS). The results of this study showed a low rate of detection of the hepatitis B virus in donated blood. NAT testing demonstrates higher sensitivity for the detection of HBV, as compare to HBsAg screening.

Current Epidemiological Features of Viral Hepatitis B and C, Tuberculosis and HIV Infection In Psychiatric Hospitals

Background. Psychiatric hospitals, where patients with immunodeficiency often do not comply with basic preventive measures, as well as receiving a wide range of medical procedures, including parenteral ones, are the institutions of high risk of socially significant infections spreading.The aim was to study the current epidemiological features of parenteral hepatitis among various categories of mentally ill patients (with pulmonary tuberculosis and HIV infection) and medical personnel in a large psychiatric hospital.Materials and methods. Serological markers of hepatitis B (HB) and hepatitis C (HC) were determined in 8352 patients and 542 employees of a large psychiatric hospital using domestic diagnostic test systems.Results. Markers of HB and HC among patients were revealed in 7.2% of persons (HB — 2.8%, HC — 3.1%, and HB+HC — 1.4%). The analysis of sex, age and social characteristics of HBVand HCV-patients was conducted. Markers of HC were significantly more common in HIV-infected patients (44.4% of individuals); the main routes of transmission of HB and HC were intravenous drug use and sexual intercourse. Among patients with pulmonary tuberculosis, the maximum number of persons was found with markers of HB (44.3%) and HB+HC (38.2%); the main clinical form of pulmonary tuberculosis was represented by the infiltrative form (60.4%); in 53,7% of cases the bacillary forms were identified that pose a serious epidemiological risk in the spread of tuberculosis in the hospital. Analysis of the social structure showed that HBV+HC+HIV and pulmonary tuberculosis are characteristics of persons with aggravated social status. The most frequent factors of infection with HBV and HCV were longterm parenteral loading and intravenous drug use. The greatest factor of parenteral load was observed in mentally ill patients diagnosed with HB+HC accompanying tuberculosis. It was shown that the frequency of HBV and HCV markers detection among medical personnel depends on the department profile, work duration, frequency and risk of contact with blood during professional activity.Conclusion. A high level of comorbidity of mental disorders and socially significant infectious diseases (HIV infection, tuberculosis and HB and HC) has been established, which has a significant impact on the epidemic process of these infections. Preventive programmes established in hospitals and in the territories they serve should take into account their comorbidity. The necessity of specific and non-specific prevention of viral hepatitis in patients and medical staff of psychiatric hospitals is shown.

Current Epidemiological Features of Viral Hepatitis B and C, Tuberculosis and HIV Infection In Psychiatric Hospitals

Background. Psychiatric hospitals, where patients with immunodeficiency often do not comply with basic preventive measures, as well as receiving a wide range of medical procedures, including parenteral ones, are the institutions of high risk of socially significant infections spreading.The aim was to study the current epidemiological features of parenteral hepatitis among various categories of mentally ill patients (with pulmonary tuberculosis and HIV infection) and medical personnel in a large psychiatric hospital.Materials and methods. Serological markers of hepatitis B (HB) and hepatitis C (HC) were determined in 8352 patients and 542 employees of a large psychiatric hospital using domestic diagnostic test systems.Results. Markers of HB and HC among patients were revealed in 7.2% of persons (HB — 2.8%, HC — 3.1%, and HB+HC — 1.4%). The analysis of sex, age and social characteristics of HBVand HCV-patients was conducted. Markers of HC were significantly more common in HIV-infected patients (44.4% of individuals); the main routes of transmission of HB and HC were intravenous drug use and sexual intercourse. Among patients with pulmonary tuberculosis, the maximum number of persons was found with markers of HB (44.3%) and HB+HC (38.2%); the main clinical form of pulmonary tuberculosis was represented by the infiltrative form (60.4%); in 53,7% of cases the bacillary forms were identified that pose a serious epidemiological risk in the spread of tuberculosis in the hospital. Analysis of the social structure showed that HBV+HC+HIV and pulmonary tuberculosis are characteristics of persons with aggravated social status. The most frequent factors of infection with HBV and HCV were longterm parenteral loading and intravenous drug use. The greatest factor of parenteral load was observed in mentally ill patients diagnosed with HB+HC accompanying tuberculosis. It was shown that the frequency of HBV and HCV markers detection among medical personnel depends on the department profile, work duration, frequency and risk of contact with blood during professional activity.Conclusion. A high level of comorbidity of mental disorders and socially significant infectious diseases (HIV infection, tuberculosis and HB and HC) has been established, which has a significant impact on the epidemic process of these infections. Preventive programmes established in hospitals and in the territories they serve should take into account their comorbidity. The necessity of specific and non-specific prevention of viral hepatitis in patients and medical staff of psychiatric hospitals is shown.

Viral Hepatitis B and C Detection among Ebola Survivors and Health Care Workers in Makeni, Sierra Leone

Viral hepatitis B and C infections are among the leading cause of death in Sub-Saharan Africa. Lack of knowledge and awareness in the general population as in health care settings may enhance the propagation of these diseases. We aimed at determining the prevalence of HBV and HCV in Ebola survivors and health care workers (HCWs) of the Makeni town in Sierra Leone. We conducted a cross-sectional study during the last 2013-2016 Ebola outbreak in Makeni among Ebola survivors (N = 68) and 81 Health care workers from Holy Spirit hospital and Loreto clinic, two health care facilities in Makeni district. Serological markers of HBV (HBs Ag, anti-HBs Ab and anti-HBc Ab) and anti-HCV antibodies detection were done using ELISA techniques. The positive detection rates for HBs Ag, anti-HBs Ab and anti-HBc antibodies in Ebola survivors were 23.53% (16/68), 32.35% (22/68) and 88.89% (16/18) respectively. Survivors with a current HBV infection had a positive rate of 38.89% (7/18) and 16.66% (3/18) of them were considered immune due to past HBV infection. HCV prevalence was 26.47% (18/68) and about 10.29% (7/68) were HBV/HCV co-infected. The positive detection rates of HBsAg, anti-HBs Ab and anti-HBc Ab were 37.07% (30/81), 33.33% (27/81) and 30.86% (25/81) respectively in health care workers. We observed that 4.94% (4/81) of the HCWs were currently infected with HBV. Participants considered as immune due to past infection represented 23.47% (19/81) and those immune due to vaccination represented 2.47% (2/81). The prevalence of HCV infection among health staff was 2.47% (2/81) with 1.23% (1/81) being HBV/HCV co-infection. Our findings showed that viral hepatitis infection is a burden for Sierra Leone government. There is an urgent need to develop and implement strategies that could improve population immunization against HBV and vulgarization of HCV treatment programs.

Variety of the hepatitis B virus genovariants in the military

Aim. To estimate the distribution of genotypes and subgenotypes of the hepatitis B virus among military personnel with chronic viral hepatitis B. Materials and methods. The work used samples of blood plasma and biopsy material obtained from 90 active or retired military personnel with chronic viral hepatitis B with various degrees of fibrosis undergoing treatment in St. Petersburg. Primary detection of HBV was carried out by isolating nucleic acids (NK) from the blood plasma using the «AmplePrime Ribo-prep» commercial kit (FBIS CRIE, Moscow). Specific primers were used for the amplification and sequencing reaction. Overlapping primer pairs were used, jointly flanking 1475 base pairs (bp) fragment, including the recommended for HBV genotyping the 1169 bp Pre-S1/Pre-S2/S. Results. Among 90 samples from patients with chronic viral hepatitis B from different regions of the Russian Federation, HBV subgenotypes are represented in the following ratios: D2 = 45.6% (n=41), D1 = 32.2% (n=29), D3 = 13.3% (n=12), A2 = 6.7% (n=6), D4 and A1 by 1.1%, respectively. The distribution of HBV subgenotypes from the North Caucasian federal district (D1 – 63.6%, D2, D3, D4, A2 – by 9.1%) was significantly different from the distribution among patients from the Central and North-Western federal districts (D1-20, 9%, D2 – 58%, D3 – 16.3%, A2 – 4.8%) (χ2=11,9приp=0,0076, df=3). Uncharacteristic for the Russian Federation subgenotypes D4 and A1, representing single imported cases. The tendency to shift the distribution of genovariants due to imports of the corresponding HBV subgenotypes from other countries, including the Central Asian countries, is discussed. Conclusion. A systematic study of the HBV isolates phylogeny provides new information about the HBV subgenotypes distribution among certain population groups, including military personnel.

J-105 Novel antiviral targets for HBV elimination

Chronic hepatitis B virus (CHB) infection continues to be a major global public health issue despite the availability of effective HBV vaccines. Approximately 257 million people worldwide are infected with CHB and complications from HBV are now a leading cause of morbidity and mortality. The number of deaths from CHB now exceed >1 million per year. In 2016, the WHO Global health sector strategy on viral hepatitis, announced new goals to achieve a 90% reduction in new cases of HBV and hepatitis C (HCV) and a 65% reduction in mortality due to HBV and HCV by 2030. To achieve the ambitious HBV targets, significant progress is needed in several key intervention areas along the continuum of HBV care including (1) improved access to HBV vaccination and drugs for the prevention of mother to child transmission; (2) more rapid and affordable diagnostics for improved screening and detection of HBV; and (3) innovative therapeutics that can result in HBV cure. Current recommended therapies for CHB include either pegylated interferon-alpha which stimulates innate immune responses against HBV or oral nucleoside reverse transcriptase inhibitor (NUC) therapies which block the maturation of HBV genome into its infectious form and thus release of infectious virus containing HBV DNA. While both therapies effectively suppress HBV DNA, they rarely result in complete loss of HBsAg, which is needed to achieve functional cure, a serological state associated with a higher remission rate after treatment cessation, and lower rates of cirrhosis and hepatocellular carcinoma. Sterilizing cure is unlikely to be achievable at the current time because of the persistence of the active transcriptional template of HBV covalently closed circular DNA (cccDNA). Recently there have been promising advances in the treatment of HBV that could potentially achieve widespread functional cure and bring us closer to achieving the ambitious WHO goals of HBV elimination. These include the development of novel therapeutics that target various stages of the HBV lifecycle (eg, HBV entry, viral replication, HBV cccDNA production, and viral protein expression) and immunotherapeutic agents, as well as innovative treatment approaches with existing HBV therapies that seek to maximize the possibility of functional cure. This goal of this talk us to provide a comprehensive overview of these new developments in HBV therapeutics, covering the following key areas: (1) current goals of treatment, and definitions of HBV “cure”; (3) the HBV life cycle, antiviral targets and investigational agents currently being developed that target virus including inhibition of viral entry, HBV surface antigen production and elimination or silencing of cccDNA; and (4) the use of novel surrogate markers (ie, quantitative HBsAg) to assess treatment response in clinical trials.

A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction

BackgroundHepatitis B virus (HBV) infection is the major public health problem worldwide. In clinical practice, serological and molecular assays are the most commonly used diagnostic methods to detect HBV infection in clinical practices.MethodsHere we present a rapid and sensitive recombinase aided amplification assay (RAA) to detect HBV at 39.0 °C for 30 min without DNA extraction from serum samples. The analytical sensitivity of RAA assay was 100 copies per reaction and showed no cross reaction with human immunodeficiency virus (HIV) and hepatitis C virus (HCV). The universality of RAA assay was validated by testing of 41 archived serum samples with predefined HBV genotypes (B, C and D).ResultsA total of 130 archived suspected HBV infected serum samples were detected by commercial qPCR with DNA extraction and RAA assay without DNA extraction (heat-treatment). Compared with qPCR assay as a reference, the RAA assay obtained 95.7% sensitivity and 100% specificity and a kappa value of 0.818.ConclusionsWe developed a rapid, convenient, highly sensitive and specific method to detect HBV without DNA extraction in clinical samples. This RAA method of HBV detection is very suitable for clinical testing.

Molecular detection and DNA Methylation of HBV in a group of Iraqi patients with Hepatocellular Carcinoma

Molecular detection and DNA Methylation of HBV in a group of Iraqi patients with Hepatocellular Carcinoma