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Integrated human diagnostics and vector control
towards OneHealth
Dr. Konstantinos Mitsakakis, et al.
University of Freiburg & Hahn-Schickard, Germany
3rd WHO Global Forum on Medical Devices, 12.05.2017, Geneva, Switzerland
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
2
www.discognosis.eu
www.diagoras.eu
www.dmc-malvec.eu
https://infravec2.eu/
Background and motivation
3
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
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)
4
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
LabDisk: Single platform – multiple use
Typical workflow in a microbiology lab is scaled down to an integrated disk-shaped cartridge
Hands-on time: <5min
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 chambers12 target pathogens simultaneously
Real-time fluorescence signal detection
Total time-to-result: 70-120 min, depending on the assay
Universal design: only variable region is primers 5
NA.2
NA.1
NA.3 NA.4
NA.5
Reference: S. Hin/B. Lopez-Jimena, et al., in preparation
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
6
Full video is available at:
https://www.youtube.com/watch?v=UvcZwOXTRuk&feature=youtu.be%20
Maurice Mutro Nigo, Bunia, DRC
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
Disk preparation steps (fabrication, reagent pre-storage)
Packaging
Fabrication of disks for validation
8
Dispatch (no cold chain)
Validation in Senegal
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Benjamin Lopez-Jimena (Univ. Stirling), Cheikh Fall (Pasteur, Dakar)
Reference: S. Hin/B. Lopez-Jimena, et al., in preparation
50 disks @ 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
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
Validation in Senegal
2. Malaria Pf
3. Malaria Pv
4. Malaria Pm
6. S. Paratyphi
7. S. pneumoniae
8. CHIKV
9. DENV1
10. DENV2
11. DENV3
12. DENV4
Waste
1. Pan-malaria
5. S. Typhi
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
Reference: S. Hin/B. Lopez-Jimena, et al., in preparation
10
8 10
9
15 disks @ Central Laboratory Khartoum
Sample matrix: whole blood
Real-time recruited patient
Pan-malaria
P. falciparum
Confirmed via blood smear test
Assay time-to-positive: 10-15 min
Laboratory tests with spiked
bacteria in 200 µL whole blood
Objective: prove “simulated”
multiple detection of bacteria
Validation in Sudan
Reference: S. Hin/B. Lopez-Jimena, et al., in preparation
11
S.pneu
moniae
S. Typhi
S. Paratyphi
Pan-
malaria
P. falciparum
EU Project DMC-MALVEC (2016-2020)
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 (2017-2021)
Expansion to arbovirus vectors
Humans get sick mosquitoes transmit
12
Reference: www.dmc-malvec.eu ; https://infravec2.eu
Conclusively
Clinical utility and diagnostic impact of the LabDisk
Synergies in a broad “ecosystem” OneHealth
13
Contact:
Email: [email protected]
Tel.: +49 1525 467 9229
Thank you for your attention!
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