Diagnostic rapide: et si nous copions la nature? Schrenzel innovation day 2014 10-29-02

1. Diagnostic rapide:
et si nous copions la nature?
Prof. Jacques Schrenzel, MD
Head of the Laboratory for Bacteriology
& of the Genomic Research Laboratory
Geneva University Hospitals, Switzerland

2. Un bref passage aux urgences...

3. Un bref passage aux urgences...
Labo de
Bactériologie

4. Un bref passage aux urgences...
48 à 72 h

5. Un bref passage aux urgences...
(Kumar A, Crit Care Med 2010)

6. Un bref passage aux urgences...

7. Un bref passage aux urgences...

9. Limitation actuelle: la culture...
Culture

11. Détection moléculaire directe ?

12. Détection moléculaire directe ?
1) Préparation

13. Détection moléculaire directe ?
1) Préparation
2) Amplification/Détection
PCR
Séquençage
ESI-MS
Microarrays

15. L’étape limitante est….
la préparation de l’échantillon
Avoir au plus vite l’ADN de la bactérie!

16. Autres méthodes?
PlexID / IRIDICA (Abbott)
Depuis 2009 au GRL (HUG)

17. Autres méthodes?
Séquenceur de 3ème génération
(Oford Nanopore Technology)
Depuis 2014 au GRL
PlexID / IRIDICA (Abbott)
Depuis 2009 au GRL (HUG)

19. Sample preparation is the key!

20. Sample Preparation Work–Flow
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”

21. Sample Preparation Work–Flow
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”
MagPhase
Technology

22. The challenge
Combination and integration of
magnetic particles and microfluidics
Homogenous suspension of
magnetic particles during target
capturing process
Enhancing the mixing of magnetic
particles and overcoming the
limitation of particle diffusion

23. Next generation Magnetic Particles
Handling Technology

24. Microfluidic Automation Process

25. Excellent linearity and reproducibility
DNA extraction from MRSA lysate and detection of
the mecA gene using real-time PCR

26. Concentration and reproducibility from
large sample volumes
140
120
100
80
60
40
20
0
E. Coli DNA capture
(10 ml sample)
1 CFU/ml 0.5 CFU/ml 0.3 CFU/ml
Yield in %

27. Sample Preparation Work–Flow
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”
FibroTrap
Technology

28. FibroTrap technology: Prologue !
Horseshoe Crab
 A “fossil age” creature with first trace
reported 540 millions years ago: more than
1000 times older than “Homo sapiens”
 Still a mysterious step during Evolution

29. FibroTrap technology: Prologue !
Injured site
 Intriguing relation to bacteria as an extract
of the horseshoe crab's lymph is used by
the pharma and medical device industries
to ensure that their products are free of
bacterial contamination (Limulus assay).
 No other test works as easily or reliably for
this purpose.
Microbe
Cascade
Reactions
Activated coagulation enzyme:
(Serpin -2)
Coagulogen
monomers
Coagulin Polymer

30. FibroTrap technology: Prologue !
Coagulation for
Bacterial Defense
Coagulation for injury
protection
-540 M
years
Injured site
Cascade
Reactions
Activated coagulation enzyme:
Thrombin
Fibrinogen
monomers
Fibrin Polymer

31. Pelleting sample preparation technology
A fast and easy way to separate viable
pathogenic agents from whole blood
(1) Get the patient sample
(2) Get the infecting agents precipitated within <20 min
(3) Lyse the pellet and recover the concentrated bacteria in a small
volume (1/50th of the sample) ready for detection

33. Pelleting sample preparation technology
Vacutainer-like technology: potential
combination with various identification methods
Cultures (agar, broth, …)
Micro-cultures
MDx (PCR, sequencing …)
Spectrometry (MALDI, ESI, Raman…)

34. Sydney Brenner
(2002 Nobel prize for Medicine)

35. Sydney Brenner
(2002 Nobel prize for Medicine)
In biotechnology,
the one -"omics"
that really counts is:
economics

36. Sydney Brenner
(2002 Nobel prize for Medicine)
In biotechnology,
the one -"omics"
that really counts is:
economics

37. Acknowledgements
 Damien BAUD
 Manuela BENTO-TANGOMO
 Eve-Julie BONETTI-SARRET
 Dr Yannick CHARRETIER
 Dr Anna-Rita CORVAGLIA
 Seydina DIENE
 Dr Sélène FERRARI
 Dr Adrien FISCHER
 PD Dr Patrice FRANCOIS
 Nadia GAIA
 Myriam GIRARD
 Dr Vladimir LAZAREVIC
www.genomic.ch
Dr Amar RIDA