1. Persistent human erythrovirus infection in blood donors National Blood Service, UK Div. Transfusion Medicine, University of Cambridge, UK Public Health Laboratory, Ankara, Turkey D Candotti, N Etiz, A Parsyan, JP Allain

2. Issues and objectives Background Chronic anaemia may occur in transfused immunosuppressed patients Some preliminary evidence of persistent B19 infection (DNA+IgG) Study objectives - to determine the prevalence of persistent B19 infection in blood donors - to characterise persistent B19 genome - to assess the potential infectivity of persistent B19

3. NAT assays Human erythrovirus genotypes (HEV 1 (NIBSC 99/800) 2 (EDQM) 3 (D91.1 plasmid) Nested-PCR 1 95% detection limit 2 Q-PCR 3 95% detection limit 2 Dynamic range 25 50 50-5x10 10 25 50 50 - 5x10 5 25 50 50 - 5x10 10 1. Primers in the NS1 region 2. IU/ml or copies/ml 3. TaqMan-based assay using primers and probe in the NS1 region

4. Human erythrovirus DNA in blood donors

5. UK South Africa Malawi References 0.01 100 83 97 73 Genotype 1 Genotype 2 Genotype 3 Ghana Phylogenetic analysis of human erythrovirus 800 nt NS1/VP1u sequences

6. Serology of HEV DNA+ samples (Biotrin IgM & IgG kits) UK Ghana Pos Neg Pos Neg IgM 1 8 1 12 IgG 9 0 10* 2 * S/COs of positives are significantly lower than with UK samples

7. Sample id: IgG status: UK4523 positive UK4892 positive Gh2094 positive Gh2120 positive Gh1280 negative Gh2135 negative 0 10 20 30 40 50 60 70 80 90 100 % of the viral population Eluted Flow through HEV DNA quantification in 6 plasmas chromatographed on protein G

8. IgG antibody to HEV genotype 1 and 3 CountryN samples Reactive to antigen (%) testedGenotype 1Genotype 3* UK 87 53 (60.9) ND Ghana 155109 (70.3) 117 (75.5) * VP2 antigen from V9 strain was provided by Dr K Brown, NIH, USA

9. Conclusions The prevalence of persistent human erythrovirus infection in random blood donors is approximately 1% Viral load in persistent HEV infections is low (median 10E3) 5-70% of persistent HEV appears uncomplexed (free) HEV Genotype 3 is prevalent in Ghana Serologic detection of antibody to genotype 3 with genotype 1 reagent is unsatisfactory

10. Acknowledgements Transfusion Medicine Unit, Komfo Anokye Teaching Hospital, Kumasi, Ghana Francis Sarkodie Shirley Owusu-Ofore National Institute for Biological Standards and Control, UK Dr. Sally Baylis Liverpool School of Tropical Medicine, UK Dr. Imelda Bates Natal Blood Transfusion Service, Pinetown, South Africa Dr. Theresa Nel Virology Lab., Armand trousseau Hospital, Paris, France Dr. Annabelle Servant Pr. Antoine Garbarg-Chenon Dept. of Microbiology, Scientific Institut of Public Health, Brussels, Belgium Dr. Isabelle Thomas Hematology branch, National Heart, Lung, and Blood Institut, Bethesda, USA Dr. Kevin Brown West Khartoum Umdurman Hospital Khartoum, Sudan Dr. A Youssef