1. Phylogenetic analysis of genotype 3 parvovirus B19 in Ghana, West Africa

2. Epidemiology Markers Anti-B19 IgG (%) Anti-B19 IgM** (%) B19 Viral DNA (%) Viral load range (IU/mL) Blood Donors 82 8 1.3 1.6x10 2 - 1.3x10 5 Children* 7.9 35 11.8 9.1x10 1 - 1.7x10 6 Pregnant Women 81 19 1.8 4.6x10 1 - 3.6x10 6 *Age range 0.13-66 months, hospitalised for severe anaemia mostly related to acute malaria (>90%). **Prevalence in viraemic B19-infected individuals.

3. Prevalence (%) of B19 DNA and anti-B19 IgG in 170 Ghanaian children according to age Age groups % prevalence 0 5 10 15 20 25 30 <12m12-23m24-3536-47>48 B19 DNA Anti-B19 IgG

4. AJ717293 AY064476 AY064475 AY903437 DQ333426 BB19 AF162273 M24682 AY386330 Z68146 M13178 DQ408301 AY504945 AY083234 GhD3-5 GhP748 GhP693 DQ408302 DQ408305 DQ408303 DQ408304 AJ249437 GhP227 GhP416 GhD1599 GhD28-8 0.005 substitutions/site 100 G1 G2 G3b G3a Phylogenetic tree of parvovirus B19 based on nearly full-length genome sequences (4866 bp)

5. AB030693 AB030673 M13178 Z70560 DQ408301 AY386330 Z68146 AF113323 BB19 Z70528 AY028237 Z70599 AY504945 M24682 AF162273 AB030694 AY044266 DQ333426 AY903437 AJ717293 DQ333428 AY064476 AY064475 DQ333427 DQ408305 DQ408302 AY083234 GhD3-5 GhP748 GhP693 DQ408304 DQ408303 GhP416 GhP227 GhD28-8 GhD1599 AJ249437 0.005 substitutions/site NS1 100 82 G1 G2 G3b G3a AB030693 AB030673 M24682 Z70528 AB030694 AF162273 AF161224 AY028237 Z70560 BB19 DQ408301 Z70599 AY504945 Z68146 AF113323 AF161226 AY386330 M13178 AF161223 AF161225 AY044266 DQ333426 AY903437 DQ333428 AJ717293 AY064476 AY064475 DQ333427 DQ408305 DQ408302 DQ408304 DQ408303 AY083234 GhD3-5 GhP693 GhP748 AY647977 GhP416 GhP227 GhD28-8 GhD1599 AJ249437 0.005 substitutions/site 95 VP1u 100 99 93 G2 G3a G1 G3b 51 100 Phylogenetic tree of parvovirus B19 based on NS1 and VP1u sequences

6. G3b AB030693 AB030673 DQ408301 Z70528 AY028237 M13178 Z70560 Z68146 AF113323 AY386330 BB19 AY504945 M24682 AF162273 Z70599 AB030694 DQ408305 DQ408302 DQ408304 DQ408303 AY083234 GhP748 GhP693 GhD3-5 GhP416 GhP227 GhD28-8 GhD1599 AJ249437 DQ333427 DQ333426 AY903437 DQ333428 AY064476 AY064475 AJ717293 0.005 substitutions/site VP2 100 G1 G2 G3a M13178 M24682 AF162273 BB19 DQ408301 AY504945 Z70528 Z68146 AY386330 Z70560 DQ408305 DQ408302 DQ408304 DQ408303 AY083234 GhD3-5 GhP693 AY647977 GhP748 GhP416 GhP227 GhD28-8 GhD1599 AJ249437 DQ333427 DQ333426 AY903437 AY064476 AY064475 0.005 substitutions/site 100 89 90 11-kDa 90 94 G1 G2 G3a G3b Phylogenetic tree of parvovirus B19 based on VP2 and 11-kDa sequences

7. Inter-(sub)group genetic diversity based on pairwise nucleotide-substitution differences (mean % distance [range]) Genome region Full genome VP2 3a vs 3b 5.42 (4.92-5.71) 6.18 (5.65-6.67) 3a vs 1 13.4 (13.2-13.7) 12.7 (12.1-13.3) 3b vs 1 13.2 (12.9-13.6) 13.0 (12.6-13.7) 3a vs 2 9.39 (9.02-9.60) 10.1 (9.49-10.8) 3b vs 2 9.32 (8.83-9.56) 10.5 (9.79-11.2)

8. Nucleotide-substitution rates * Mean rate of nucleotide substitutions per site per year. † Shackelton LA & Holmes EC. 2006. J Virol 80:3666-69. HPD, 95% high-probability-density intervals; TMRCA, times from the most recent common ancestor. Dataset B19 genotype 3 NS1+VP1 B19 genotype 1† VP1 Mean* (HPD) 2.0x10 -4 (2.0x10 -6 ; 5.1x10 -4 ) 1.14x10 -4 (1.20x10 -5 ; 2.40x10 -4 ) TMRCA 525 (43.5; 1992) - Clock Strict

9. 0.001 substitutions/site Phylogenetic tree based on NS1/VP1u sequences of B19 genotype 3 strains from Ghana (N=53), Western Europe (N=10), and Brazil (N=7)

10. Conclusions Two genetically distinct clusters identified within B19 genotype 3 (>5% mean genetic diversity) Classification of genotype 3 strains into two subtypes (B19/3a and B19/3b) is proposed Potential West African origin of B19 genotype 3 Similar rate of evolutionary change of B19 genotype 3 strains than those of B19 genotype 1 and carnivore parvoviruses High prevalence of persistent B19 genotype 3 infection (~1.4%) - no evidence of maternofetal transmission - no evidence of transfusion-transmitted infection

11. Acknowledgements Dr D. Candotti National Blood Service Cambridge, UK Pr J.-P. Allain Dr A. Parsyan Div. of Transfusion Medicine University of Cambridge Cambridge, UK Dr S. Owusu-Ofori Mr F. Sarkodie Transfusion Medicine Unit Komfo Anokye Teaching Hospital Kumasi, Ghana Dr K. Danso Dr E. Addo-Yobo Ms H. Akpene Dept. of Obstetrics & Gynaecology Komfo Anokye Teaching Hospital Kumasi, Ghana Mr A. Dompreh Dept. of Microbiology Komfo Anokye Teaching Hospital Kumasi, Ghana Dr C. Szmaragd Dept. of Genetics University of Cambridge Cambridge, UK Dr N. Etiz Refik Saydam Hygiene Centre Dept. of Virology Ankara, Turkey Dr S. Kerr Dr G. Elliott Biotrin Ltd. Dublin, Ireland