1. Detection of Dengue Virus RNA in Blood Donors from Honduras and Brazil with a Prototype Transcription-Mediated Amplification Assay

2. Dengue Over 2.5 billion people live in risk areas for dengue infection WHO estimates that in 2004 there were 50-100 million cases of DF, 500,000 cases of dengue hemorrhagic shock syndrome (DHS) and 20,000 consequent deaths Epidemic is expanding due to global warming, expanding vectors and trafficking of subtypes Dengue the most important arboviral disease of humans

3. Expanding global epidemic of Dengue

4. Number of dengue fever cases reported per month, Brazil. A) 1986–1993, B) 1994–2003. Dark bars represent January.

5. Curvas epidémicas del 2002, 2003,2004 Y 2005* Casos Sospechosos de Dengue Clásico Honduras, *Semana 41 * AÑO EPIDÉMICO 1995 EXCLUÍDO 2003 (16,559 casos) FUENTE: Telegrama Epidemiológico. 2002 (32,269 casos) 2004 (19,971 casos) 2005 (16,037 casos)*

6. Analytical sensitivity of the prototype dengue TMA assay DEN 1, 2, 3 and 4 diluted to 1 PFU/mL

7. Sources of Donor Plasma from Dengue Endemic Regions CountryHondurasBrasilAustralia RegionTequciqalpa San Pedro Sula Sao Paulo Cairin s Townsville Brisban e Cairins & Townsville Period 08/31/04- 01/24/05 09/06/04- 01/13/05 Feb. 2003- Apr. 2003 2003 2005 Total by Region 1187181932393220107114442500 Total by Country 299432398235 12 IR, 9 RR 11 confirmed 8 IR, 3 IR confirmation pending in progress

8. Dengue TMA Results from Honduran Blood Donors (n = 2994) Specimen ID Initial TMA Results Repeat TMA Results BSRI Genotyping CDC Genotypin g Growth in culture (CDC) Duplicate Viral Load Testing (PFU/100 uL of plasma) DV-00033Reactive Nonreactiv e DEN-1Negative <0.001 DV-00164Reactive DEN-4 0.44 DV-00220Reactive DEN-4 13.07 DV-00477Reactive DEN-4 51.84 DV-00545Reactive DEN-1 1.38 DV-00766Reactive Nonreactiv e Negative Neg DV-00802Reactive DEN-4Negative 0.25 DV-01110Reactive DEN-1 Negative0.54 DV-01328Reactive DEN-1 Negative20.00 DV-02221Reactive Nonreactiv e DEN-3Negative 0.002 DV-02837Reactive DEN-2 Negative0.01 DV-02921Reactive DEN-3Negative 0.002

9. Planned studies of TT-Dengue Transmission study in Honduras with BSF and Gen-Probe funding Transmission study in Brazil under REDS- II funding Collaborations with Australia, Hong Kong and Singapore blood banks

10. Dengue Transmission Study Approach It is mandatory in Brazil to save donor serum samples for 6 months after the transfusion Phase 1: Prevalence Study –Dengue TMA on 10,000 unlinked donor samples to determine prevalence Phase 2: Transmission Study –collect recipient samples ~1 week after transfusion. –capture donor samples in routine repository. –Dengue TMA on both samples. –sequence virus to confirm linkage

11. Acknowledgements J Linnen, A Broulik, C Collins, DP Kolk, C Giachetti, L Mimms Gen-Probe Incorporated, San Diego, CA; E Vinelli Honduran Red Cross, Tegucigalpa, Honduras; E Sabino, Blood Center of Sao Paulo, Sao Paulo, Brazil; R Lanciotti, CDC, Fort Collins, CO; C Hyland, Australian Red Cross, Brisbane, Australia; L Tobler, MP Busch Blood Systems Research Institute, San Francisco, CA;