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

Epidemiology Markers Anti-B19 IgG (%) Anti-B19 IgM** (%) B19 Viral DNA (%) Viral load range (IU/mL) Blood Donors x x10 5 Children* x x10 6 Pregnant Women x x10 6 *Age range months, hospitalised for severe anaemia mostly related to acute malaria (>90%). **Prevalence in viraemic B19-infected individuals.

Prevalence (%) of B19 DNA and anti-B19 IgG in 170 Ghanaian children according to age Age groups % prevalence <12m12-23m >48 B19 DNA Anti-B19 IgG

AJ AY AY AY DQ BB19 AF M24682 AY Z68146 M13178 DQ AY AY GhD3-5 GhP748 GhP693 DQ DQ DQ DQ AJ GhP227 GhP416 GhD1599 GhD substitutions/site 100 G1 G2 G3b G3a Phylogenetic tree of parvovirus B19 based on nearly full-length genome sequences (4866 bp)

AB AB M13178 Z70560 DQ AY Z68146 AF BB19 Z70528 AY Z70599 AY M24682 AF AB AY DQ AY AJ DQ AY AY DQ DQ DQ AY GhD3-5 GhP748 GhP693 DQ DQ GhP416 GhP227 GhD28-8 GhD1599 AJ substitutions/site NS G1 G2 G3b G3a AB AB M24682 Z70528 AB AF AF AY Z70560 BB19 DQ Z70599 AY Z68146 AF AF AY M13178 AF AF AY DQ AY DQ AJ AY AY DQ DQ DQ DQ DQ AY GhD3-5 GhP693 GhP748 AY GhP416 GhP227 GhD28-8 GhD1599 AJ substitutions/site 95 VP1u G2 G3a G1 G3b Phylogenetic tree of parvovirus B19 based on NS1 and VP1u sequences

G3b AB AB DQ Z70528 AY M13178 Z70560 Z68146 AF AY BB19 AY M24682 AF Z70599 AB DQ DQ DQ DQ AY GhP748 GhP693 GhD3-5 GhP416 GhP227 GhD28-8 GhD1599 AJ DQ DQ AY DQ AY AY AJ substitutions/site VP2 100 G1 G2 G3a M13178 M24682 AF BB19 DQ AY Z70528 Z68146 AY Z70560 DQ DQ DQ DQ AY GhD3-5 GhP693 AY GhP748 GhP416 GhP227 GhD28-8 GhD1599 AJ DQ DQ AY AY AY substitutions/site kDa G1 G2 G3a G3b Phylogenetic tree of parvovirus B19 based on VP2 and 11-kDa sequences

Inter-(sub)group genetic diversity based on pairwise nucleotide-substitution differences (mean % distance [range]) Genome region Full genome VP2 3a vs 3b 5.42 ( ) 6.18 ( ) 3a vs ( ) 12.7 ( ) 3b vs ( ) 13.0 ( ) 3a vs ( ) 10.1 ( ) 3b vs ( ) 10.5 ( )

Nucleotide-substitution rates * Mean rate of nucleotide substitutions per site per year. † Shackelton LA & Holmes EC J Virol 80: 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

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)

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

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