Differential diagnosis of fever in West- and East-Africa S. Hin1, B. Lopez-Jimena3, M.A. Bakheit4, V. Klein2, S. Stack5, C. Fall6, A.A. Sall6, K. Enan7, S. Frischmann4, L. Gillies5, M. Weidmann3, S. Goethel8, V. Rusu8, O. Strohmeier1,2, N. Paust1,2, R. Zengerle1,2, K. Mitsakakis1,2,* 1 Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg, Germany 2 Hahn-Schickard, Freiburg, Germany; 3 Institute of Aquaculture, University of Stirling, United Kingdom 4 MAST Diagnostica GmbH, Reinfeld, Germany; 5 Mast Group Limited, Liverpool, United Kingdom 6 Arbovirus and viral haemorrhagic fever unit, Institut Pasteur de Dakar, Dakar, Senegal 7 Central Laboratory, Khartoum, Sudan; 8 MagnaMedics Diagnostics BV, Geleen, The Netherlands * E-mail: konstantinos.mitsakakis@imtek.de Tel: +49 761 203 73252 Summary The FeverDisk point-of-care test detects on a single disposable cartridge and in a fully automated way, up to 12 fever-causing pathogens (viruses, bacteria, parasites) enabling, proper patient management and epidemics surveillance. L Motivation Acute fever is one of the most common symptoms among patients globally Diagnosis based merely on clinical symptoms is highly risky and inaccurate [1] Co-infections and epidemics outbreaks complicate the diagnostic landscape, often leading to wrong diagnosis and treatment [2] Proposed solution The FeverDisk as a point-of-care diagnostic tool [3] Results 50 disks @ Institut Pasteur de Dakar, Senegal [3] 15 disks @ Central Laboratory Khartoum, Sudan [3] Biobanked & fresh samples were used Conclusions Successful demonstration in operative environments Co-infections were detected (malaria/chikungunya & dengue/chikungunya) Unknown dengue serotype in sample was identified Positive and negative results on disk were confirmed by (RT)-qPCR, microscopy, lateral flow tests Capacity building of local personnel Acknowledgements / Disclosure This work was supported by the EC project DiscoGnosis (www.discognosis.eu) (FP7 GA-318408). We thank the Hahn-Schickard Lab-on-a-Chip Foundry Service for the production of the LabDisks. No conflict of interests is applicable. References [1] D’Acremont, V., et al., New England Journal of Medicine, 370, 2014, pp.809-817. [2] M. Amexo, et al., Lancet, 364, 2004, pp.1896-1898. [3] S. Hin/B. Lopez-Jimena, et al., in preparation. (A) (B) (C) Figure 3: (A) Packaged LabDisks ready for use. (B) Dr. Cheikh Fall using the LabDisk & Player at the Institut Pasteur de Dakar (March 2016. Source. Dr. B. Lopez-Jimena) Pan-malaria Chikungunya P. falciparum Dengue-2 Key parameters Performance features Multiplexed panel 12 targets simultaneously Sample volume 200 µL (blood, serum) Sample preparation In situ, all reagents for DNA/RNA extraction & purification are pre-stored Hands-on time < 5 min Total time-to-answer 70-120 min depending on the assay Amplification DNA/RNA: LAMP isothermal (primers and enzymes pre-stored) Stability studies Accelerated shelf life studies at > 90% humidity indicate 1.6 year stability @ 22oC Figure 4: Detection of double infection in biobanked sample in Dakar, Senegal. No unspecific amplification observed. Sample matrix: serum Figure 5: Detection of malaria in a real-time recruited patient in Central Laboratory Khartoum, Sudan. Sample matrix: whole blood NA.2 NA.3 NA.4 NA.1 Workflow step-by-step Sample addition in NA.1 Extraction and purification of pathogens’ DNA/RNA using pre-stored buffers (NA.2) Mixing with amplification enzymes (NA.3) Distribution to discrete reaction chambers (pre-stored primers offer specificity, NA.4) Isothermal amplification and real-time fluorescence signal detection (NA.4) Figure 1: The LabDisk used in the study Diagnostic panel of the FeverDisk (adaptable to end-users’ needs) Bacteria S. Typhi S. Paratyphi S. pneumoniae Parasites Pan malaria screening P. falciparum P. malariae P. vivax Viruses Dengue (1-4) Chikungunya Zika Febrile patient & antimicrobial resistance management Early epidemics identification & surveillance Vector control, disease data management Figure 2: Clinical utility and diagnostic impact of the FeverDisk