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

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www.discognosis.eu

www.diagoras.eu

www.dmc-malvec.eu

https://infravec2.eu/

Background and motivation

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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)

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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

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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

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Disk preparation steps (fabrication, reagent pre-storage)

Packaging

Fabrication of disks for validation

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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

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8 10

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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

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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

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Reference: www.dmc-malvec.eu ; https://infravec2.eu

Conclusively

Clinical utility and diagnostic impact of the LabDisk

Synergies in a broad “ecosystem” OneHealth

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Contact:

Email: konstantinos.mitsakakis@imtek.de

Tel.: +49 1525 467 9229

Thank you for your attention!

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