1. HBV genotypes E and A deletions and recombination: two sides of the same coin? Penelope Garmiri 1, Andre Loua 2, Daniel Candotti 3 and Jean-Pierre Allain 1 1 Dept of Haematology, University of Cambridge 2 National Blood Transfusion Centre, Conakry, Guinea 3 NHSBT, Cambridge

2. HBV genotype E became prevalent in West Africa over the past 300 years HBsAg carriage ranging between 12% and 25% is the highest in the world In Guinea, the prevalence of HBsAg+ in 1st time blood donors is 15% Very high HBV exposure in population (75%) HBV prevalence in Guinea, West Africa

3. Phylogenetic analysis of full genome sequences from Guinea 81 full genome sequences Genotype A3 : 1 seq Genotype E : 77 seq Seq in blue: Recombinants

4. 7 Guinean strains with deletions in the core region A 3222 bpEcoRI Pre-core Core 18182454 Wild type  1 1979 2241 262bp  2 23142007 49bp147bp  4 20102235 78bp37 bp B

5. Recombinant strains Indication – Differential clustering of sequences during assembling – Unusual clustering of samples during phylogenetic analysis Confirmation – NCBI/ Genotyping function – SIMPLOT software

6. Guinean recombinant strains GU1616, GU754, GU763: full genome GU1125, GU1127, GU1607: partial sequences GH16, GH2537: full genome from Ghana Further confirmation - Cloning

7. Evidence of recombination between HBV genotype E and A and E and D in strains originating from Guinea and Ghana Red: genotype A Blue: genotype E Orange: genotype D

8. Schematic representation of Guinean and Ghanaian HBV A/E and E/D recombinant strains

9. Schematic representation of clone diversity from a single plasma sample from Ghana (GH99)

10. SIMPLOT analysis of GU1127 recombinant clones

11. SIMPLOT analysis of GU1607 recombinant clones

12. SIMPLOT analysis of GU1616 recombinant clones

13. Overview of the recombination and deletion points in West African strains Deletions and recombinations occur in similar core regions suggesting preferential DNA re-arrangement in this area of HBV genome

14. Viral Load distribution of HBsAg+ Guinean samples

15. The ratio E/A3 is 77/1 but 7% of donors carry recombinant forms (RFs) Recombination points are identical in multiple strains - Recombination occurs in the same positions of the HBV genome and/or - RFs are infectious (CRFs) It most likely happened earlier when the balance of E and A genotypes in the population was different Viral Load of Rec samples is similarly distributed to the WT samples Origin & epidemiology of Rec HBV in Guinea

16. Summary Genotype E is most prevalent in Guinea (93.9%) Core deletions in Guinea strains (8.5%); evidence of multiple populations Recombination events between genotypes E and A are frequent in Guinea (7.1%) Several E/A infectious recombinant forms might be circulating in the region Deletions and recombinations occur in the core region suggesting preferential DNA rearrangement in this area of HBV genome

17. Acknowledgements The staff of National Blood Transfusion Centre, Conakry for collecting and testing the donor samples Dr I.H.A. Barnes for determining the HBV BCP/PC sequences of the strains included in this study Dr B. Meldal for her input in the phylogenetic analysis Funding from National Health Service Blood and Transplant (NHSBT)