HBV Infection In Blood Donors Research Articles

Plasma Cytokine Profile in Occult HBV-Infected Blood Donors

Aim: Cytokine profile in occult HBV infection (OBI) was systematically investigated to identify the immunopathogenesis of OBI. Materials & methods: A total of 46 OBI, ten asymptomatic hepatitis B surface antigen carriers, ten chronic hepatitis B and 12 healthy blood donors were recruited. A total of 21 plasma cytokines were detected. Results: Compared with healthy blood donors, elevated plasma Th1, Th2, Th17 and immune regulatory associated cytokines were observed in OBI. Almost no significant difference was found for these cytokines among OBI, asymptomatic hepatitis B surface antigen carriers and chronic hepatitis B. OBI displayed the predominance of type 2 and regulatory immunity. Conclusion: OBI displayed the general cytokine profile of chronic HBV infection, which might contribute to virus persistence and the presence of the liver microinflammatory environment. The clinical implications of OBI deserve more attention.

Comparative analysis of hepatitis B virus infections in blood donors born before and after the implementation of universal HBV vaccination in southern China.

In China, the vaccinated blood donors have rapidly increased by recent years, which may impact blood safety. The true prevalence of HBV between vaccinated blood donors and non-vaccinated blood donors should be explored. The samples of blood donors were collected and detected for serologic markers of HBV in the Shenzhen Blood Centre (SZBC). The discrepant results were tested with commercial electrochemiluminescence immunoassay (ELCI) for HBsAg, anti-HBs, HBeAg, Anti-HBe and Anti-HBc, alternative MPX ID NAT, nested PCR, and a quantitative real-time polymerase chain reaction (qPCR) assay for HBV DNA. The serological and molecular characteristics of HBV infected blood donors were analysed, and the effects on blood safety for donors born before and after the implementation of universal HBV vaccination were compared. Out of 242 presumed HBV infected donors from 26 318 donations, 131 (0.49%, [95% CI, 0.43-0.59]) chronic HBV infections (CHB, HBsAg detected with or without DNA), 58 (0.22%, [95% CI, 0.17-0.28]) occult hepatitis B infections (OBI, HBsAg not detected, assume anti-HBc positive and/or anti-HBs with HBV DNA) and 3 (0.011%, [95% CI, 0.0023-0.033]) window period (WP) infections were confirmed respectively. There were 28 CHBs (0.44%), 7 OBIs (0.11%) and 1 WP (0.016%) from vaccinated blood donor and 103 CHBs (0.52%), 51 OBIs (0.26%) and 2 WPs (0.01%) from non-vaccinated blood donor. The HBV+ (CHBs, OBIs and WPs) rate (0.56%) in vaccinated donors was lower than in non-vaccinated donors (0.78%, p < 0.05). The HBsAg titers of vaccinated infected blood donors (Median: 128.8IU/ml) were much higher than non-vaccinated infected blood donors (58.4IU/ml). The OBI yield rates in the vaccinated blood donors was significantly lower than the non-vaccinated blood donors (p < 0.05). There 102/124 (82.3%) samples were genotype B, 22/124 (17.7%) were genotype C respectively. There was no significant difference in the distribution of genotype between non-vaccinated blood donors (B/C, 86/17) and vaccinated blood donors (B/C, 23/6; p > 0.05). High frequency of vaccine escape mutations M133L (32.4%) and E164G in S region of genotype B strains and substitution L175S (40.9%) related to vaccine escape in S region of genotype C strains were identified. The universal HBV vaccination program markedly reduces the risk of HBV infection in blood donors, and provides a significant guarantee for the safety of blood transfusion. Several important mutations detected related vaccine escape and notable mutations needed further investigated.

Study on Reentry Evaluation Mode for Blood Donors Used to be HBV Reactive in Jiangsu Province

To evaluate the risk of reentry in HBV reactive blood donors and feasibility of HBV reentry strategy. HBsAg+ or HBV DNA+ donors who had been quarantined for more than 6 months in Jiangsu Province could propose for reentry application. Blood samples were routinely screened by dual-ELISA for HBsAg, anti-HCV, HIV Ab/Ag, and anti- Treponema pallidum and those non-reactive ones were tested by minipool nucleic acid testing (NAT) for three times. To identify occult HBV donors, samples of NAT non-reactive were further tested by electrochemiluminescence immunoassay (ECLIA) for HBV seromarkers (including HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb). Donors of only 4 ECLIA patterns were accepted to reentry, including all 5 HBV seromarkers negative, anti-HBs only but having history of hepatitis B vaccine injection, HBcAb only, HBsAb+ / HBcAb+ with HBsAb more than 200 IU/L. Additionally, the detection rate of HBV infection was compared between routine screening mode and ECLIA, as well as the reentry qualified rate of HBsAg+ and HBV DNA+ blood donors. From Oct. 2016 to Aug. 2019, a total of 737 HBV reactive donors had applied for reentry, including 667 HBsAg+ reactive and 70 HBV DNA+ reactive donors. Among 3 screening methods, the highest HBV detection rate (43.15%, 318/737) was observed on ECLIA, while only 4.75% (35/737) on ELISA and 3.12% (23/737) on NAT, respectively. Among 4 qualified patterns of HBV serological markers, the highest proportion was found in the all negative group (22.90%, 155/677), followed by the group with HBsAb+ only and history of hepatitis B vaccine injection (19.35%, 131/677), and the median concentration of HBsAb was 237.7 IU/L. The unqualified rate of HBV DNA+ donors was 82.86%, which was significantly higher than 47.98% of HBsAg+ donors. Routine screening tests merely based on ELISA and NAT could miss occult HBV donors and may not be sufficient for blood safety. HBsAb concentration and vaccine injection history should be included in the evaluation of HBV reactive donors who intend to apply for reentry. There is a relatively larger residual risk of occult HBV infection in blood donors quarantined for HBV DNA reactive.

Metavirome composition of Brazilian blood donors positive for the routinely tested blood-borne infections

Viral metagenomics is widely applied to characterize emerging viral pathogens but it can also reveal the virome composition in health and disease. The evaluation of the virome in healthy blood donors can provide important knowledge on possible transfusion threats. Currently, there is still a paucity of information regarding the virome of blood donors who test positive for routinely tested blood-borne infections. Such analysis may reveal co-infections which in turn appear to be crucial for transfusion medicine and for patient management. The aim of this study was to evaluate the metavirome in blood donors who tested positive for routinely tested blood-borne infections, the information for which is important for transfusion medicine and blood donor management. For this purpose, we analyzed 18 blood donations obtained from HIV and HBV-infected blood donors from the Brazilian Amazon (Amapa state) and 11 HIV, HBV, HCV, syphilis and Chagas disease - positive blood donations obtained from blood donors sampled in South Brazil (Rio Grande do Sul state). We additionally included a control group of 20 blood donors obtained from Southeast Brazil (State of São Paulo). Samples were assembled in pools and sequenced by the Illumina NovaSeq 6000 platform. To link a given virus with geographic region or type of blood donor, we performed supervised machine learning classification (fingerprint analysis). The virome of both locations was predominantly composed of commensal viruses. However, in HBV-infected blood donors from the Brazilian Amazon, the Human Pegivirus-1 (HPgV-1) reads were prevailing, while in HIV-infected donors from the same location, the torque teno virus (TTV) reads expressive abundance. In blood donors from South Brazil, the most abundant reads were classified as Human endogenous retrovirus K (HERV-K). Putative emerging viruses like the Human gemykibivirus-2 (HuGkV-2) were exclusively identified in samples from the Brazilian Amazon. The fingerprint analysis demonstrated that the HERV-K, TTV-7, 13, and 15 were statistically important for the infected blood donors, while TTV-5, 12 and 20 were linked to geographic localization. Our study revealed differences in the viral composition among blood donors who tested positive for routinely tested blood-borne infections from two different Brazilian regions and indicated the presence of putative emerging viruses in samples obtained from the Amazon. Together our results show that the presence of specific commensal viruses may be related donor infection status but additional investigations including larger study groups and samples from other Brazilian regions are needed to confirm this hypothesis.

Assessment of hepatitis B surface antigen negative blood units for HBV DNA among replacement blood donors in a hospital based blood bank in Nigeria.

Hepatitis B virus infection is one of the greatest threats to blood safety all over the world. The laboratory algorithm based on only the detection of hepatitis B surface antigen (HBsAg) leaves a gap for infected HBsAg negative donors to donate blood during the "window period" (WP) and late stages of infection. To estimate the frequency of the presence of HBV deoxyribonucleic acid (DNA) in HBsAg negative blood units screened using two different assays for HBsAg in a high endemic region. Frozen serum aliquot of 100 replacement blood donors who donated blood units that were HBsAg negative were retrieved and tested for HBV DNA. Sample positive for HBV DNA was sequenced by Sanger's method, genotyped and the viral load was determined. One sample (1%) was positive for HBV DNA. The HBV viral load of the sample was 768,000 IU/ml. The partial S-gene of the Hepatitis B virus isolated was genotype E using the NCBI viral genotyping tool. There is still a risk of HBV infected blood unit escaping detection when donor testing is limited to HBsAg screening. The use of NAT which can substantially reduce HBV infected blood donors from the donor pool should be considered.

Hepatitis B infections among blood donors in England between 2009 and 2018: Is an occult hepatitis B infection a risk for blood safety?

Hepatitis B virus (HBV) is one of the most frequent infections identified in blood donors in England and represents an ongoing blood safety risk. We have analyzed markers of HBV infections in blood donors in England between 2009 and 2018 and used these to estimate the likelihood of non-detection of occult HBV infection (OBI). We collected epidemiological, virological, and genotyping information on HBV cases identified in England, 2009-2018. The estimated risk of non-detection and likely transmission of OBI were compared to lookback and transfusion-transmitted infections surveillance data. Six-hundered and fifty-five HBV-infected blood donors were identified in England during the 10-year period; 598 chronic, 32 acute, and 25 occult HBV infections. However, most donors with chronic and occult infections were born in Eastern Europe, Africa, or Asia (451/544, 83% and 14/24, 58%); acute infections were largely seen in UK-born donors (19/28, 68%). Genotyping of 266 HBV-positive samples revealed five genotypes (A-E), reflecting ethnicity and country of birth. Most OBIs were identified in repeat donors (19/25); lookback data identified a transmission rate of 8.3%. It is estimated that at least 13 potentially infectious donations from donors with OBI remain undetected annually, equating to an overall residual transmission risk of 3.1 per million donations using our current screening strategy of HBsAg screening with HBV nucleic acid testing (NAT) in pools of 24. OBI accounted for the majority of the HBV residual risk in England. Further cost-benefit analysis is required to estimate if our current HBV screening strategy should be changed.

Serological Patterns of Hepstitis B Virus Among HBsAg Negative Blood Donors in Baghdad, Iraq by Using Architect System

Background: About 10-15% of population HBV infection becomes chronic, and 85-90% are resolved. In Iraq, HBs-Ag assay by ELISA is only mandatory test for detection of HBV infection in blood donors. Anti- HBc and anti-HBs were not detectable in their sera.Objective: To investigate the distribution of serological patterns for HBV who were HBsAg negative.Material and method: Between July 18, 2011 to December 25, 2011, 10ml of blood samples were collected from National blood Transfusion Center donors, Baghdad, for 25782 individuals (25294 male, 488 female), mean age (20-65)years. HbsAg positive sera excluded (185), and final number of donors included in thestudy was 25597 HbsAg negative.Result: The data showed a previous exposure for HBV(p&gt;0.05) through anti-HBc and anti-HBs 3.8% by Architect system. The anti-HBs combine with anti-HBc was 68.5%, while anti-HBc alone was 31.5%. Independent risk factors were association with married, history of blood transfusion and residence.Conclusion: The prevalence of the HBV in Iraq is 4.6% as intermediate region at first time.

Molecular evaluation of HBV core gene mutations in asymptomatic HBV infected blood donors in Iran.

Mutations in the core promoter and precore regions of HBV cause down-regulation of HBeAg. These mutations are associated with chronic hepatitis, cirrhosis and Hepato Cellular Carcinoma (HCC). This study was carried out to sequence analysis of HBV core gene in HBsAg- positive blood donors in Iran. A total of 50 HBsAg- positive blood donor samples were examined in this study. Serological markers of hepatitis B including: HBsAg, HBeAg, HBeAb and HBcAb were measured by ELISA method. HBV-DNA was extracted from the sera, and then PCR was performed on extracted HBV-DNA using specific primer of gene C. After direct sequencing, the nucleotide sequences from 50 blood donors were analyzed using a reference sequences and then phylogenetic analysis was performed. Also, the line probe assay was used to detect mutations. The majority of donors (62.5%) were in the age group of 29 - 40 years old. Among all the HBV DNA positive cases, 87.8% were HBeAg negative. The prevalence of PC and BCP mutants were 12% and 55% respectively, among asymptomatic HBV infected blood donors by direct sequencing method. The results of this study showed that some of HBV infected blood donors had mutation in core gene of HBV and amino acid changes in B cell, T helper and CTL epitopes that can cause reducing HBe and HBc antigenicity in asymptomatic HBV infected blood donors and the development of escape mutants from host immune.

Increasing prevalence of HDV/HBV infection over 15 years in France

BackgroundIn France, there are no consistent data estimating hepatitis delta virus (HDV) prevalence in the general population. ObjectivesTo better characterize HDV/HBV infection and its trends over a 15-years period from 1997 to 2011, we used data retrieved from the National Epidemiological Donors database including viral and demographic characteristics of all French HBV infected blood donors. Study designOf the 39,911,011 donations collected over the 15 year-study-period, 6214 (1.56 in 104 donations) were confirmed positive for HBV from which 72.3% were tested for HDV antibodies (Ab). HDV viral load was performed using a real-time PCR assay on positive HDV Ab samples and HDV genotype determined for each positive viremic sample. ResultsAmong the 4492 HBV donations, 89 (1.98%) were HDV Ab positive. After being stable around 1.1% from 1997 to 2005, this rate has continuously increased to reach 6.5% in 2010, before declining to 0.85% in 2011. Of the 61 investigated HDV Ab positive individuals, 22.9% were viremic with a viral load ranging from 104 to 9.8×107copiesmL−1. Genotyping revealed 12 HDV-1, 1 HDV-6 and 1 HDV-7 in accordance with the geographical origin of individuals. ConclusionSuch a study gives unexpected features of HBV–HDV infection in the population of blood donors which is a priori, a healthy population. The increase of HDV prevalence mainly linked to migration of population from endemic countries, demonstrates that there is still no complete control of HBV infection and must encourage HBV vaccination campaigns and systematic screening for HDV in HBV-infected.

The diversity of chronic hepatitis B virus infections within blood donors in England and North Wales 2005 through 2010

In 2010 hepatitis B virus (HBV) was the most frequently detected infection in UK blood donation screening, typically found in first-time, male, chronically infected donors born abroad. To date there has been no comprehensive characterization of the virologic profile of these infections. Epidemiologic and serologic data were collected retrospectively for 344 chronically HBV-infected blood donors identified from July 2005 to June 2010. Additional laboratory testing was carried out to determine the HBV genotype, viral load, and prevalence of clinically significant mutations and to detect hepatitis delta virus (HDV) coinfection. Five HBV genotypes (A-E) were found, Genotypes D (45%), A (20%), and E (20%) were the most prevalent. A strong association was seen between genotype and donor ethnicity (p < 0.001) and between genotype and place of residence (p = 0.006). Clinically significant mutations were observed across hepatitis B surface antigen (17%), basal core promoter (25%) and precore (78%) regions. An antiviral resistance profile was identified in one donor. Evidence of HDV coinfection was found in 2% of donors. The data show the diversity of HBV in asymptomatic chronic infections detected in blood donors in England and North Wales and demonstrates the presence of mutations which may impact on disease. The global nature of these infections and the inability to identify chronically infected donors before donation highlights the importance of using screening assays capable of detecting a broad range of genotypes and mutations. Furthermore, the integration of the virologic and demographic data allows us to more accurately construct a profile of our chronically HBV-infected blood donors.

Molecular epidemiological study of hepatitis B virus in blood donors from five Chinese blood centers

Although the genetic variability of hepatitis B virus (HBV) in HBV-infected patients has been extensively studied, reports on genotypes, subtypes and mutations in the S region of HBV strains from Chinese blood donors are limited. In this study, 245 blood samples from HBsAg-positive blood donors were collected from five geographically diverse blood centers in China. The S region of HBV was amplified, and the HBV genotype and subtype were determined. The amino acid sequences of the S region were aligned, and mutations related to the failure of immunization and HBsAg detection were determined. Of the 245 samples, 228 (93 %) were genotyped successfully. We found that genotypes B, C, D and A accounted for 58.8 %, 21.9 %, 6.6 % and 3.95 % of the isolates, respectively. The distribution of HBV antigen subtypes was as follows: adw (67.6 %), adr (23.3 %) and ayw (8.7 %). Mutations were present in 39 (17.1 %) of 228 samples in the major hydrophilic region (MHR) of the S region. This study demonstrated that HBV genotype/subtype B/adw was the most frequent strain circulating in HBV-infected Chinese blood donors, followed by C/adr. The occurrence of MHR mutants in HBV-infected blood donors and the potential failure to detect some of them in collected units poses a threat to transfusion safety.

Sequence analysis of the polymerase gene in hepatitis B virus infected blood donors in Iran.

Hepatitis B is one of the most common infectious diseases worldwide that can be transmitted by blood transfusion. The hepatitis B virus (HBV) has eight different genotypes that show different geographical distributions and clinical manifestations. This study aims to investigate the sequence of the HBV polymerase gene and the frequency of HBV genotypes among Iranian blood donors. The sera of 223 blood donors who were positive for hepatitis B surface antigen (HBsAg) as determined by the ELISA method were selected. HBV DNA was extracted from the sera of 134 blood donors by a commercial kit, and the entire polymerase gene was amplified by nested-PCR. HBV genotypes were determined by direct sequencing of the HBV polymerase gene. Phylogenetic trees were constructed by the neighbor-joining (NJ) method. No known base mutations were found in the entire HBV polymerase gene of infected blood donors, and only genotype D was detected among HBV-infected blood donors. The sub-genotype D1 of HBV was dominant in the subjects. This study shows that antiviral-resistant mutations, such as lamivudine-resistant HBV strains, do not exist naturally among Iranian blood donors. More studies on the full-length HBV genomes are required to determine genome evolution of HBV among infected Iranian blood donors.

Nucleic Acid Testing to Detect HBV Infection in Blood Donors

Nucleic Acid Testing to Detect HBV Infection in Blood Donors

Nucleic Acid Testing to Detect HBV Infection in Blood Donors

Nucleic Acid Testing to Detect HBV Infection in Blood Donors

Frequency of hepatitis B virus DNA in anti-HBc positive, HBsAg negative blood donors in Rasht, northern Iran

Background One of the important factors in the ensuing safety of blood transfusion is to use a sensitive screening assay for detection of blood-born infective agents such as HBV which transmits through transfusion. To improve the detection rate of HBV infection in blood donors, a cross-sectional study was conducted in Rasht, which is the largest city in the north of Iran to explore the possibility of using anti-HBc as a screening test. Study design and methods A total of 2041 blood samples negative for HBsAg, Anti-HCV, Anti-HIV I, II and RPR were tested to detect anti-HBc and then the positive anti-HBc samples were further checked for the presence of HBV DNA. Results The prevalence of anti-HBc positive samples was 3.8% and HBV DNA was detected in only one sample. Conclusions This study showed that anti-HBc positive blood donors may be a source of HBV transmission and further study for evaluation of HBV DNA in anti-HBc positive blood units is needed.

The Relationship between HBSAG and HBV DNA in HBV-Infected Blood Donors

The Relationship between HBSAG and HBV DNA in HBV-Infected Blood Donors

HBsAg non-reactive HBV infection in blood donors: Transmission and pathogenicity

Five blood donors with occult persistent and one donor with an early window phase HBV infection were identified. They were negative in regular HBsAg screening and had low levels of HBV DNA that were probably not detectable by current mini-pool nucleic acid amplification testing. In four donors several mutations were found located in the HBs antigen loop. In three donors the mutations were predominantly outside the “a” determinant; one donor had a wild type HBsAg sequence. Fifty-five recipients of donations from the persistently infected donors could be tested for previous or ongoing HBV infection and of them 53% (29/55) were anti-HBc positive. Based on the prevalence of anti-HBc in Germany (7%), it was assumed that four of those recipients had already been positive before transfusion. In 22 cases, it was assumed that they acquired infection by the donations, but the infection remained asymptomatic and was resolved. In three cases transmission was proven by the time course of the acute infection and sequence identity The resulting infection was fatal and associated with immunological disorders at the time of transmission: in one case sepsis and in the other two cases immunosuppression. In a further asymptomatic case of proven transmission from the early window phase donation passively administered anti-HBs could not prevent spread of wildtype HBV but antiviral treatment lead to resolution. Surprisingly the typical acute hepatitis B was not observed in a single one of the 26 cases of assumed or proven transmission. J. Med. Virol. 79:S32–S36, 2007. © 2007 Wiley-Liss, Inc.

Transfusion-transmitted hepatitis B virus infection in the UK: a small and moving target.

Transfusion-transmitted hepatitis B virus (TT-HBV) infections, when analysed in detail provide information about the nature and relative frequency of the sources of infectious donations. These cases are therefore used to inform blood safety strategies. This study updates previous reviews of the causes of TT-HBV in order to determine whether a change may have occurred in recent years. Cases of TT-HBV reported during 1998-2001 were reviewed and the nature of the infectious donations described. These cases were compared to a previously published case series reported during 1991-97. Six cases of TT-HBV have been reported in the UK between 1998 and 2001. All were the result of infectious donations collected from donors with acute HBV infection. This is in contrast to the series reported during 1991-97 when only three of 14 similar cases were caused by acute infections in donors, with the majority of incidents being the result of chronic infection in donors. There appears to have been a change in the relative importance of acute and chronic HBV infection in blood donors in causing TT-HBV infections. Improvements in the sensitivity of HBsAg assays and/or a decrease in the prevalence of chronic HBV infection in blood donors could explain this observation. This change may have implications for strategies to reduce the risk of TT-HBV infection.

The precore/core promoter mutant (T1762A1764) of hepatitis B virus: clinical significance and an easy method for detection.

Recently, a new hepatitis B virus (HBV) mutant with HBe antigen-negative phenotype has been characterized, in which one TATA box-like motif of the precore/core promoter had degenerated: most frequently by both A-->T and G-->A mutations at positions 1762 and 1764, respectively. The clinical significance of this mutant is as yet unknown. In our present study, the T1762 A1764 mutant was sought in sera from HBV-infected blood donors and chronic liver disease patients by directly sequencing a PCR-amplified region of HBVDNA. Also, because the A1764 mutation generates a Sau3AI cleavage site (GGTC-->GATC), we digested the PCR products with Sau3AI to see if cleavage would occur at this specific site. Our results mostly corroborated the earlier report but we found a higher-than-predicted frequency of HBe antigen-positive blood donors positive for the mutant (22%). The titres of HBe antigen in these mutant-positive sera were slightly decreased compared to the titres in wild-type HBV infection. In addition, these blood donors had relatively high (though within the normal range) serum alanine aminotransferase (ALT) levels, suggesting that the T1762 A1764 mutation could be used as a sensitive laboratory marker for insidious hepatitis in these otherwise 'asymptomatic' carriers. The Sau3AI assay, which is much more convenient than sequencing, was shown to be useful for the detection of the T1762 A1764 mutant in an extensive number of clinical samples.