Molecular characteristics of HBV infection among blood donors tested HBsAg reactive in a single ELISA test in southern China

Abstract Background Hepatitis B virus (HBV) infection is a major concern for blood safety in high-prevalence HBV countries such as China. In Shenzhen, dual hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assays (ELISAs) have been adopted in parallel with nucleic acid testing (NAT) for donors for over a decade. A small proportion of blood donors test reactive (R) for HBsAg but negative through routine NAT, which can lead to HBV infection with an extremely low viral load. Objectives We aimed to investigate and analyze the molecular characteristics of HBV among blood donors that tested HBsAg R in a single ELISA test. Methods Blood donations were evaluated in this study if confirmed HBsAg R through one of two ELISA kits. Samples with non-reactive (NR) results by NAT were collected and tested for HBsAg by chemiluminescent microparticle immunoassay (CLIA) with a neutralization test. The level of HBsAg was further assessed by electrochemiluminescence immunoassay (ECLIA). The viral basic core promoter (BCP) and pre-core (PC) and S regions were amplified by nested PCR. Quantitative real-time PCR (qPCR) for viral load determination and individual donation (ID)-NAT were adopted simultaneously. HBsAg was confirmed with CLIA, ECLIA, nested PCR, qPCR, and ID-NAT. Results Of the 100,252 donations, 38 and 41 were identified as HBsAg R with Wantai and DiaSorin ELISA kits, respectively. Seventy-nine (0.077%, 79/100,252) blood samples with ELISA R-NR and NAT NR results were enrolled in the study. Of these, 17 (21.5%,17/79) were confirmed as HBsAg-positive. Of the 14 genotyped cases, 78.6% (11/14) were genotype B, and C and D were observed in two and one sample, respectively. Mutations were found in the S gene, including Y100C, Y103I, G145R, and L175S, which can affect the detection of HBsAg. A high-frequency mutation, T1719G (93.3%), was detected in the BCP/PC region, which reduced the viral replication. Conclusion A small number of blood samples with HBsAg ELISA R-NR and NAT NR results were confirmed as HBV infection, viral nucleic acids were found in most of the samples through routine NAT methods. It is necessary to employ more sensitive and specific assays for the detection of HBV infection among blood donors.

Download Full-text

Molecular Characteristics of HBV Among Blood Donors in Southern China with Different HBsAg ELISA Results by Two Kits

10.21203/rs.3.rs-76662/v1 ◽

2020 ◽

Author(s):

Xianlin Ye ◽

Tong Li ◽

Ran Li ◽

Heng Liu ◽

Junpeng Zhao ◽

...

Keyword(s):

Viral Load ◽

Hepatitis B ◽

Blood Donors ◽

Southern China ◽

Core Promoter ◽

Hepatitis B Surface Antigen ◽

Hbv Infection ◽

Molecular Characteristics ◽

Blood Samples ◽

Elisa Kit

Abstract Background: The blood donors tested reactive (R) for hepatitis B surface antigen (HBsAg) but were missed by routine nucleic acid test (NAT), which can be resulted by the infection of hepatitis B virus (HBV) with extremely low viral load. This phenomenon remains a strict threat to blood transfusion in China.Objectives: We aimed to investigate and analyze the molecular characteristics of HBV among blood donors that detected as HBsAg reactive (R) in single ELISA Reagent. Methods: The blood donations were detected as HBsAg R in one ELISA kit, using two kinds of ELISA reagents. These samples with non-reactive (NR) results by NAT were collected and tested for HBsAg by Abbott chemiluminescent microparticle immunoassay (CLIA) with neutralization test, the level of HBsAg were further detected by Roche electrochemiluminescence immunoassay (ECLIA). The viral basic core promoter (BCP) and pre-core (PC) and S regions were also amplified by nested-PCRs. Quantitative real-time PCR (qPCR) for viral load determination and individual donation NAT of Roche reagent were adopted simultaneously. HBsAg was confirmed by the results of CLIA, ECLIA, nested-PCR, qPCR, and ID-NAT.Results: Of 100,252 donations, 38 and 41 were identified as HBsAg reactive only in WanTai and DiaSorin ELISA kit respectively, after blood screening using WanTai and DiaSorin ELISA assays. 79 (0.077%, 79/100,252) blood samples with ELISA R-NR and NAT NR results were enrolled in the study. Of which, 17 (21.5%,17/79) were confirmed as HBsAg positive. Of 14 cases genotyped, genotype B were 78.6% (11/14), C and D was observed in 2 (14.2%, 2/14) and 1 sample (7.1%, 1/14), respectively. Mutations in the S gene such as Y100C, Y103I, G145R, and L175S were found, which can affect the detection of HBsAg. A high-frequency mutation, T1719G (93.3%), was detected in BCP/PC and would reduce the replication of the virus. Conclusion: A small part of blood samples with HBsAg ELISA R-NR and NAT NR results were confirmed as HBV infection, viral nucleic acids were found in most of those samples through the in-house NAT methods. It is necessary to apply more sensitive and specific assays for the detection of HBV infection among blood donors.

Download Full-text

Molecular characteristics of chronic hepatitis B virus infection among voluntary blood donors in Kinshasa, Democratic Republic of the Congo

10.1101/740803 ◽

2019 ◽

Author(s):

Pati Moloko Maindo ◽

Didier Anzenza Mudwahefa ◽

Jean-Pierre Kambala Mukendi ◽

Sylvain Ramazani Yuma ◽

Jérémie Masidi Muwonga ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B ◽

Blood Donors ◽

Hbv Infection ◽

Molecular Characteristics ◽

Chronic Hbv Infection ◽

Chronic Hepatitis B Virus ◽

B Virus ◽

Chronic Hbv ◽

Genotype A

AbstractBackgroundHepatitis B represents a major global health problem. Despite the high endemicity of hepatitis B in Sub-Saharan Africa, little is known about the molecular characteristics of chronic hepatitis B virus (HBV) infection in Africa, and there are very few published studies that describe the genetic characteristics of HBV in asymptomatic adults in DRC. The present study aimed at determining the molecular diversity of chronic HBV infection in voluntary blood donors in Kinshasa, DRC.MethodsBlood samples from 582 voluntary blood donors at the National Blood Transfusion Centre in Kinshasa, DRC, were screened for hepatitis B surface antigen (HBsAg) using enzyme-linked immunosorbent assay (ELISA). Partial amplification and sequencing of S gene in HBV-positive samples was conducted.ResultsThe presence of HBsAg was detected in 6.9 % (40/582) blood donors. Phylogenetic analysis based on partial S gene nucleotide sequences of HBV showed that the majority (66.7 %, 10/15) of HBV strains clustered into genotype A, followed by the genotypes E (26.6 %, 4/15) and D (6.7 %, 1/15). Genotype A strains were classified into subgenotype A1, quasi-subgenotype A3, and subgenotype A4, with quasi-subgenotype A3 being predominant. One new genotype A strain did not cluster with any existing HBV/A subgenotype or quasi-subgenotype.ConclusionsThe present study highlights the high genetic variability of chronic HBV infection in DRC, and the possibility of a new HBV/A subgenotype, suggesting that HBV has a long evolutionary history in DRC. Further molecular characterization of complete HBV genomes is needed for a more accurate assessment of HBV genetic variability and its clinical significance in DRC, as partial sequences are not appropriate for determining HBV new subgenotypes.

Download Full-text

SAT0140 SAFETY OF TOFACITINIB THERAPY IN HBSAG CARRIERS WITH RHEUMATOID ARTHRITIS: A PROSPECTIVE STUDY

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.6353 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1008.2-1008

Author(s):

L. Fang ◽

Z. Lin ◽

Z. Liao ◽

O. Jin ◽

Y. Pan ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Viral Load ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Hbv Infection ◽

Hbsag Carriers ◽

B Virus ◽

Group 2 ◽

A Prospective Study ◽

Group 1

Background:Targeted synthetic DMARDs (ts-DMARDs) are becoming more available and affordable in developing countries, where the prevalence of hepatitis B virus (HBV) infection is still an important public health issue. The safety of ts-DMARDs therapy in terms of the reactivation of hepatitis B virus (HBV) infection need more concern. Rare data from a prospective study focus on the use of ts-DMARDs in patients with concurrent rheumatoid arthritis (RA) and HBV infection were available by now.Objectives:To evaluate the influence of tofacitinib on reactivation of HBV infection in HBsAg carriers with RA.Methods:In this 52 weeks observation, HBsAg carriers with active RA (DAS28>5.1) despite failed combined treatment with MTX and other non-biological DMARDs were enrolled. Patients must have normal liver function prior to study. All patients received therapy with tofacitinib (5mg twice daily) and concomitant MTX (10-12.5mg/w). Entecavir was prescribed preventively for patients who had a baseline HBV load >2000 copy/ml (group 1), and Lamivudin for patients with HBV load ≤ 2000 copy/ml (group 2). Liver enzymes (AST/ALT) and HBV viral load were monitored every 4 weeks. Increased viral load and abnormal liver function were managed according to expert opinion.Results:Thirteen patients (10 female) were recruited. Nine patients had a baseline viral load >2000 copy/ml (group 1, with preventive Entecavir), and the other 4 patients had a viral load ≤ 2000 copy/ml (group 2, with preventive Lamivudin). Two patients from group 1 discontinued tofacitinib at week 12 due to ineffectiveness, and both continued taking Entecavir for another 3 months after the discontinuation of tofacitinib.No reactivation of hepatitis B was observed in patients from group 1. One patients (female, 54 years old) from group 2 underwent a mild increase of both ALT and AST (67 and 56 IU/L, respectively) at week 16. An elevated viral load (4.9e6 copies/ml, baseline 1.4e3) and a HBV YMDD mutant was also found. The tofacitinib treatment continued. After prescription of Adefovir (combined with the pre-existing Lamivudin), both liver enzyme and viral load decreased to normal range in 8 weeks and remained normal throughout the study.Conclusion:An aggressive Tofacitinib + MTX therapy may be a safe option for HBsAg carriers with cs-DMARDs refractory RA. More active and effective prophylaxis strategy may be recommended to reduce the risk of HBV reactivation during the treatment.References:[1]Chen YM, Huang WN, Wu YD, et al. Reactivation of hepatitis B virus infection in patients with rheumatoid arthritis receiving tofacitinib: a real-world study. Ann Rheum Dis 2018; 77:780-2.Disclosure of Interests: :None declared

Download Full-text

Determination of hepatitis B virus DNA incidence, viral load, and mutations in blood donors with HBsAg and anti-HBs-negative serology and antibodies to hepatitis B core antigen

European Journal of Internal Medicine ◽

10.1016/j.ejim.2007.07.001 ◽

2007 ◽

Vol 18 (8) ◽

pp. 571-575 ◽

Cited By ~ 12

Author(s):

Duygu Findik ◽

Ugur Arslan ◽

Mahmut Baykan

Keyword(s):

Viral Load ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Blood Donors ◽

Core Antigen ◽

Hepatitis B Virus Dna ◽

B Virus ◽

Hepatitis B Core Antigen

Download Full-text

Seroprevalence of Hepatitis E Virus Infection among Blood Donors in Bulgaria

Viruses ◽

10.3390/v13030492 ◽

2021 ◽

Vol 13 (3) ◽

pp. 492

Author(s):

Magdalena Baymakova ◽

Krasimira Terzieva ◽

Rumen Popov ◽

Elisaveta Grancharova ◽

Todor Kundurzhiev ◽

...

Keyword(s):

Virus Infection ◽

Hepatitis E Virus ◽

Blood Donors ◽

Hepatitis E ◽

Elisa Test ◽

Blood Samples ◽

Wild Boars ◽

Domestic Pigs ◽

Specific Questionnaire ◽

Positive Results

Hepatitis E virus (HEV) infection is widespread among domestic pigs, industrial swine, and wild boars in Bulgaria. The aim of the current research was to present the HEV seroprevalence among blood donors in Bulgaria. In the present study, 555 blood donors (479 males and 76 females) were enrolled from five districts in the country (Shumen, Pleven, Stara Zagora, Plovdiv, and Sofia districts). All blood samples were tested for anti-HEV IgG using the recomWell HEV IgG ELISA test (Mikrogen GmbH, Neuried, Germany). Each participating donor completed a short, structured, and specific questionnaire to document data on the current study. Anti-HEV IgG positive results were detected in 144 (25.9%) blood donors, including 129 (26.9%) males and 15 (19.7%) females. The established HEV seropositivity was 28.8% (23/80) in Shumen district, 23.2% (22/95) in Pleven district, 27.1% (38/140) in Stara Zagora district, 27.5% (44/160) in Plovdiv district, and 21.3% (17/80) in Sofia district. A high HEV seroprevalence was found for persons who declared that they were general hunters (48.7%; 19/39; p = 0.001) and hunters of wild boars (51.6%; 16/31; p = 0.001). We present the first seroprevalence rates of HEV infection in blood donors from Bulgaria. The results of our research showed high HEV seropositivity among blood donors.

Download Full-text

HIV and Hepatitis B seroprevalence among the nepalese blood donors

Journal of Pathology of Nepal ◽

10.3126/jpn.v3i5.7864 ◽

2013 ◽

Vol 3 (5) ◽

pp. 390-393 ◽

Cited By ~ 1

Author(s):

SR Kandel ◽

P Ghimire ◽

BR Tiwari ◽

M Rajkarnikar

Keyword(s):

Hepatitis B ◽

Blood Donors ◽

Hepatitis B Surface Antigen ◽

Elisa Test ◽

Age Group ◽

Male And Female ◽

Test Kit ◽

The Difference ◽

Donated Blood ◽

Donor Recruitment

Background: HIV and Hepatitis B infections are public health problems in Nepal. This study was conducted based at NRCS/CBTS, with the objective of determining the HIV and HBsAg sero-prevalence in non-remunerated volunteer blood donors. Materials and Methods: A total of 66,904 units of blood collected, following donor recruitment criteriaduring March 2009-Sept. 2010 was included for analysis. All donated blood samples were subjected to screening for Transfusion transmitted infections including HIV and Hepatitis B surface antigen using standard ELISA test kits (Dade Behring, Germany). Initial reactive sera were re-tested for reconfi rmation with same test kits plus another test kit (Detect-HIV, Adaltis Inc, and Qualisa). Results: Out of 66,904 units of blood collected, 56,973 units were from male and 9,931 were from female donors. Among the total screened samples, 73 (0.10%) were found to be positive for HIV, {0.11% (64/56973) in male and 0.09% (9/9931) in female}; the difference between male and female donors (?2<3.841) was statistically signifi cant. The seroprevalence of HIV was highest in age group of 30- 39 both in male and female (p<0.001). Similarly, for HBsAg, overall seroprevalence was found to be 0.47% (316/66904 {0.42% (242/56973) in male and 0.74% (74/9931) in female}. The difference was statistically signifi cant (?2<3.841). The highest HBsAg sero-prevalence(0.65%) was also observed in same age group i.e. 30-39 (p<0.001) in male but highest seroprevalence (2.63%) was observed inage group of ?50 in female. Conclusion: Both HIV and HBV sero-prevalence is high in adult voluntary blood donors. Journal of Pathology of Nepal (2013) Vol. 3, No.1, Issue 5, 390-393 DOI: http://dx.doi.org/10.3126/jpn.v3i5.7864

Download Full-text

Molecular Diagnosis in Differentiating Active and Inactive Forms of Hepatitis B Virus Carriers

Journal of University of Babylon for Pure and Applied Sciences ◽

10.29196/jubpas.v26i10.1838 ◽

2018 ◽

Vol 26 (10) ◽

pp. 41-45

Author(s):

Salah Tofik Jalal Balaky ◽

Saeed Ghulam Hussain ◽

Amer Ali Khaleel ◽

Furat Tahseen Sabeer ◽

Ahang Hasan Mawlood

Keyword(s):

Nucleic Acid ◽

Hepatitis B ◽

Cross Sectional Study ◽

Hbv Dna ◽

Hbv Infection ◽

Detection Methods ◽

Rt Pcr ◽

Cross Sectional ◽

Blood Samples ◽

Acid Test

Background & objectives: Introducing a nucleic acid test program is aimed to diagnose and reduces the risk of viral infection or transmission. DNA assay for HBV can detect infection in the windows period, chronic occult infection and can discriminate between active and inactive HBV infection. This cross-sectional study designed to diagnose, analyze HBV infection and to differentiate active from inactive infection based on viral DNA detection. Methods: Blood samples were collected from 256 patients previously diagnosed on the clinical ground as hepatitis B seropositive in Erbil Central Lab. The viral nucleic acid quantitative assessment was done for the collected samples using RT-PCR. Q-square was performed for statistical analysis. Results: Out of 256 collected blood samples 93 (36.3%) showed HBV-DNA positive titers above 50 IU/ml. Among positive subjects, 67 (72.04%) was categorized as inactive carriers (˂ 2000-20.000 IU/ml HBV-DNA titers). Conclusions: The data produced from this study confirmed the importance of the RT-PCR technique in sensitivity and reliability as a superior diagnostics tool specifically in differentiating active from inactive HBV carriers.

Download Full-text

Age-wise and Gender-wise Prevalence of Hepatitis B Virus (HBV) Infection in Lahore, Pakistan

BioScientific Review ◽

10.32350/bsr.0104.03 ◽

2019 ◽

Vol 01 (04) ◽

pp. 20-28

Author(s):

Aqib Nazeer ◽

Shahid Ali ◽

Imran Tipu

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Hepatitis B Surface Antigen ◽

Hbv Infection ◽

Age Group ◽

Pakistani Population ◽

Blood Samples ◽

Significant Difference ◽

B Virus ◽

And Gender

Background The prevalence of hepatitis B virus (HBV) in the Pakistani population has been reported previously, however, studies with a city-oriented approach and focus on age and gender distribution are very limited. Therefore, the current study was designed to unravel the age-wise and gender wise prevalence of HBV in Lahore, Pakistan. Methods A total of 350 blood samples of both male and female patients who visited National Genetic Laboratory, Lahore between February 2019 and July 2019 and who were suspected of HBV infection were screened. Sandwich based ELISA was used to detect rapid hepatitis B surface antigen (HbsAg) according to the manufacturer’s instruction. Real time PCR was used to detect HBV using HBV Rotor Gene PCR kit. Results Out of 350 blood samples screened for HBV infection (n= 350), 180 (51.43%) were of males and 170 (48.57%) were of females. Mean age (years) with SD (standard deviation) of the screened population was 37.22 ± 12.16 years. Overall, 224 samples (64%) were found to be positive for HBV infection. In our study, the number of females with this infection (52.24%) was slightly higher than males (47.76%). However, we observed no statistically significant difference (p = 0.225) between them. Conclusion Our study concludes that HBV is highly prevalent in Lahore, Pakistan. Females are slightly more susceptible to HBV infection as compared to males. This study also reports that HBV is more prevalent in the 20-40 age group.

Download Full-text

The risk of hepatitis B virus infection by transfusion in Kumasi, Ghana

Blood ◽

10.1182/blood-2002-04-1084 ◽

2003 ◽

Vol 101 (6) ◽

pp. 2419-2425 ◽

Cited By ~ 102

Author(s):

Jean-Pierre Allain ◽

Daniel Candotti ◽

Kate Soldan ◽

Francis Sarkodie ◽

Bruce Phelps ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Blood Donors ◽

Hepatitis B Surface Antigen ◽

Hbv Dna ◽

Hbv Infection ◽

Sub Saharan Africa ◽

B Virus ◽

Sub Saharan ◽

Hbv Transmission

The risk of hepatitis B virus (HBV) transmission by transfusion in sub-Saharan Africa is considered to be relatively low, and testing of blood donors is often not done or is done relatively poorly. To re-examine this attitude, we identified HBV chronically infected blood donors from a major hospital in Ghana with a range of hepatitis B surface antigen (HBsAg) assays. Test efficacy was estimated using HBV DNA as a gold standard, and the risk of HBV infection in blood recipients was estimated for different testing strategies. Particle agglutination, dipstick, and enzyme immunoassay (EIA) HBsAg screening detected 54%, 71%, and 97% of HBV infectious donors, respectively. The risk of HBV transmission to recipients less than 10 years old ranged between 1:11 and 1:326 with blood unscreened and screened by EIA, respectively. For older recipients, the risk decreased a further 4-fold because of the high frequency of natural exposure to HBV. A total of 98% of HBsAg-confirmed positive samples contained HBV DNA. HBV DNA load was less than 1 × 104 IU/mL in 75% of HBsAg-reactive samples, most of them anti-HBe reactive. Approximately 0.5% of HBsAg-negative but anti-HBc-positive samples contained HBV DNA. The use of sensitive HBsAg tests is critical to prevent transfusion transmission of HBV infection to young children in a population with a 15% prevalence of chronic HBV infection in blood donors. However, this will not have much effect on the prevalence of this infection unless other strategies to protect children from infection are also advanced in parallel.

Download Full-text