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Molecular characterization of hepatitis B virus in blood donors in Botswana

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Abstract

Hepatitis B virus (HBV) poses a significant threat to blood transfusion safety in sub-Saharan Africa (SSA) where allogeneic blood donations are screened serologically, and more sensitive nucleic acid tests (NATs) are utilized infrequently. HBV strains circulating among blood donors in Botswana are not yet characterized. We designed a cross-sectional study to determine the HBV sub-genotypes and prevalence of hepatitis B surface antigen (HBsAg) among blood donors between November 2014 and October 2015. A total of 12,575 blood donations were screened for HBsAg and 50 consecutive plasma samples were selected for genotyping from confirmed HBsAg+ donations. Overlapping Pol and complete S (Pol/S) open reading frames (ORFs) were sequenced from extracted HBV DNA. To identify any signature amino acids, mutations were compared to sequences from a cohort of chronic HBV patients co-infected with HIV and were treatment naïve. The prevalence of HBsAg+ blood donors was 1.02% (95% CI 0.9–1.2%), and the circulating sub-genotypes were A1 serotype adw2 (36.1%), D2 serotype ayw2 (2.9%), and D3 serotypes ayw 1/2 (58.3%). Prevalence of escape mutations was 14% from HBV isolates of blood donors and 15% from isolates of HBV/HIV co-infected patients (p = 0.6926). The escape mutations sP120L, sG130R, sY134H, and sD144A were identified predominantly among HBV isolates from blood donors. These escape mutations have been associated with accelerated HBV sequelae [e.g., liver cirrhosis (LC) and hepatocellular carcinoma (HCC)], failure to detect HBsAg, inability to respond to immunoglobulin (Ig) therapy, and HBV vaccine escape. Characterizing the HBV burden, circulating sub-genotypes, and clinically relevant mutations among blood donors in Botswana is important to elucidate the efficacy of currently available vaccines, predicting HBV-transmission patterns, understanding the cohort’s risk to HBV-related complications, and to developing prevention strategies and effective genotype-based antiretroviral therapies.

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Acknowledgements

The authors would like to acknowledge the support and collaborations from National Blood Transfusions Services of Botswana. We also acknowledge the Ministry of Health and Wellness of Botswana for granting the permissions to conduct this pioneer study for molecular epidemiology of HBV in blood donors in Botswana. We would like to also extend our acknowledgements to the National University of Science and Technology, Zimbabwe; University of Botswana; and the Botswana-Harvard HIV Reference Laboratory for their sponsorship, support, and contribution to the success of the study.

Funding

This work was supported through the Sub-Saharan African Network for TB/HIV Research Excellence (SANTHE), a DELTAS Africa Initiative [Grant No. DEL-15-006]. The DELTAS Africa Initiative is an independent funding scheme of the African Academy of Sciences (AAS)’s Alliance for Accelerating Excellence in Science in Africa (AESA) and supported by the New Partnership for Africa’s Development Planning and Coordinating Agency (NEPAD Agency) with funding from the Wellcome Trust [Grant No. 107752/Z/15/Z] and the UK government. The views expressed in this publication are those of the author(s) and not necessarily those of AAS, NEPAD Agency, Wellcome Trust, or the UK government.

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Authors and Affiliations

  1. Research Laboratory, Botswana Harvard AIDS Institute Partnership, Gaborone, Botswana

    Wonderful T. Choga, Motswedi Anderson, Sikhulile Moyo, Tshepiso Mbangiwa, Bonolo B. Phinius, Pinkie Melamu, Max Essex, Rosemary M. Musonda & Simani Gaseitsiwe

  2. Department of Applied Biology and Biochemistry, National University of Science and Technology, Bulawayo, Zimbabwe

    Wonderful T. Choga & Edward Zumbika

  3. Department of Biological Sciences, University of Botswana, Gaborone, Botswana

    Motswedi Anderson & Theresa K. Sebunya

  4. Department of Medical Laboratory Sciences, Faculty of Health Sciences, University of Botswana, Gaborone, Botswana

    Tshepiso Mbangiwa & Ishmael Kasvosve

  5. National Health Laboratory (NHL), Ministry of Health and Wellness, Gaborone, Botswana

    Mukendi K. Kayembe

  6. Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA, USA

    Sikhulile Moyo, Max Essex, Rosemary M. Musonda & Simani Gaseitsiwe

  7. University of Cincinnati College of Medicine, Cincinnati, OH, USA

    Jason T. Blackard

  8. Research Laboratory, Botswana Harvard AIDS Institute Partnership, Private Bag BO 320, Gaborone, Botswana

    Simani Gaseitsiwe

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  1. Wonderful T. Choga
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  2. Motswedi Anderson
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  14. Simani Gaseitsiwe
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Contributions

WTC wrote the first draft of the manuscript. WTC and MA collaborated in the lab work, primary data analysis. SM and PM were involved in statistical analysis. TM and BBP were involved in conducting some of the lab work and manuscript editing before submission. EZ, TKS, and IK supervised the project and edited the manuscript prior to submission and data evaluation. MKK provided with samples and their demographics, and also edited the manuscript before submission. JTB developed the secondary analysis plan and performed the phylogenetics of the study. ME and RMM edited the manuscript prior to submission and complemented it with contextual data. SG developed the study, main directions of the analysis plan, overall interpretation of results and edited manuscript before submission. All co-authors read and authorized the final manuscript.

Corresponding author

Correspondence to Simani Gaseitsiwe.

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The authors declare that they have no conflict of interest.

Ethical approval

The participants’ samples used in the study were residual from blood donors, obtained already de-identified and anonymous. These were a sub-population of HBV + screened samples from National Blood Transfusion services in Botswana (NBTS). The study was approved by the University of Botswana Institute Review Board and the Health Research Development Division (HRDD) at the Botswana Ministry of Health and Wellness (HPDME 13/18/1).

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Choga, W.T., Anderson, M., Zumbika, E. et al. Molecular characterization of hepatitis B virus in blood donors in Botswana. Virus Genes 55, 33–42 (2019). https://doi.org/10.1007/s11262-018-1610-z

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