Integrated human diagnostics and vector control towards OneHealth

Transcription

1 Integrated human diagnostics and vector control towards OneHealth Dr. Konstantinos Mitsakakis, et al. University of Freiburg & Hahn-Schickard, Germany 3 rd WHO Global Forum on Medical Devices, , Geneva, Switzerland

2 Disclosure / Declaration of interests Research support has been granted by the European Commission and National Agencies via public funding LifeAssay Diagnostics Ltd (South Africa) subcontracts Hahn-Schickard in the project Antimonia funded by the Angloamerican and the South African Medical Research Council There is no conflict of interests in the presented work

3 Background and motivation Scenario: A patient presents with acute fever (especially around tropics) By default, malaria is assumed and tested accordingly Malaria is negative (non-malaria febrile illness) then what??? Additional tests? Additional costs & precious time And are they available? No? Travel to the closest clinic Will the doctor give any (available) drug acting under pressure? Acute fever is the same symptom in many diseases Overlapping epidemics complicted diagnostic landscape False diagnosis wrong therapy die / develop resistances 3

4 Goal and objectives Differential diagnosis of acute fever (febrile syndrome) via multiplex panel identification of pathogen DNA/RNA Guidance to suitable treatment management (antibiotic, anti-malarial, other...) Enhanced epidemics control and surveillance Current disease panel (all have same clinical symptoms but different treatment needs) Proving that the system works with 3 major types of pathogens, it becomes adaptable/tailored to other panels (endemic, epidemic) according to end-users needs 4

5 LabDisk: Single platform multiple use Typical workflow in a microbiology lab is scaled down to an integrated disk-shaped cartridge Hands-on time: <5min NA.3 Insert 200 µl sample (blood, serum,...) in NA.1 Lysis, extraction, purification of pathogens DNA/RNA Buffers pre-stored in NA.2 Bead-based process in NA.3 Mixing with amplification enzymes lyopellet in NA.4 Amplification in chambers with pre-stored primers (NA.5) LAMP isothermal 12 chambers 12 target pathogens simultaneously Real-time fluorescence signal detection Total time-to-result: min, depending on the assay Universal design: only variable region is primers Reference: S. Hin/B. Lopez-Jimena, et al., in preparation 5 NA.2 NA.1 NA.4 NA.5

6 The LabDisk Player Disk processing device (early prototype) Compact, portable: 26 cm x 17 cm x 10 cm; 2 kg Developed by Partner in cooperation with Hahn-Schickard Hardware Mechanical unit Thermal unit (thermocycling, isothermal) Detection unit (real-time fluorescence) One device - all disk types and applications Full video is available at: Maurice Mutro Nigo, Bunia, DRC 6

7 The LabDisk Player Disk processing device (early prototype) Compact, portable: 26 cm x 17 cm x 10 cm; 2 kg Developed by Partner in cooperation with Hahn-Schickard Hardware Mechanical unit Thermal unit (thermocycling, isothermal) Detection unit (real-time fluorescence) One device - all disk types and applications 7

8 Fabrication of disks for validation Disk preparation steps (fabrication, reagent pre-storage) Dispatch (no cold chain) Packaging 8

9 Validation in Senegal 50 Institut Pasteur de Dakar, Senegal Biobanked samples Objectives Proof-of-principle & demonstration on site Comparison with reference methods (RDTs, microscopy, (RT)-qPCR, LAMP in tube) Usability assessment Capacity building The LabDisk Player stays at Dakar Staff was trained: video-tutorial for sustainable training Benjamin Lopez-Jimena (Univ. Stirling), Cheikh Fall (Pasteur, Dakar) Reference: S. Hin/B. Lopez-Jimena, et al., in preparation 9

10 Validation in Senegal Indicative outcomes (the highlights ) In DENV samples of unknown serotype, we identified serotype 1. Sample: supernatant of infected cells In a known positive CHIKV (8) sample, we discovered DENV2 co-infection (10). Sample: serum Primers: Uni Stirling, Mast Diagnostica GmbH Lyopellets: Mast Grp Ltd Extraction buffers: MangaMedics Diagnostics BV Magn. beads: Analytik Jena The panel: different primers per chamber 1. Pan-malaria 2. Malaria Pf 3. Malaria Pv 4. Malaria Pm 5. S. Typhi 6. S. Paratyphi 7. S. pneumoniae 8. CHIKV 9. DENV1 10. DENV2 11. DENV3 12. DENV4 Waste Reference: S. Hin/B. Lopez-Jimena, et al., in preparation 10

11 Validation in Sudan 15 Central Laboratory Khartoum Sample matrix: whole blood Real-time recruited patient Pan-malaria P. falciparum Confirmed via blood smear test Assay time-to-positive: min Panmalaria P. falciparum Laboratory tests with spiked bacteria in 200 µl whole blood Objective: prove simulated multiple detection of bacteria S.pneu moniae S. Typhi S. Paratyphi Reference: S. Hin/B. Lopez-Jimena, et al., in preparation 11

12 Humans get sick mosquitoes transmit EU Project DMC-MALVEC ( ) LabDisk for molecular characterization of malaria-infected mosquitoes (species ID, parasite identification, resistance to insecticides) Disease Data Management System (DDMS): collect information from LabDisk to enhance decision support The Serious Game : interactive ICT platform for learning of workflows, communication and interpretation of data Validation in 4 African countries Côte d Ivoire, Cameroon, Ethiopia, Zambia EU project Infravec2 ( ) Expansion to arbovirus vectors Reference: ; 12

13 Conclusively Clinical utility and diagnostic impact of the LabDisk We have the tools, we invite for partnerships Contact: Tel.: Climate & environmental changes Therapeutics & vaccines Veterinaryzoonotic diseases Thank you for your attention! Synergies in a broad ecosystem OneHealth 13