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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
Un bref passage aux urgences...
Labo de
Bactériologie
Un bref passage aux urgences...
48 à 72 h
Un bref passage aux urgences...
Un bref passage aux urgences...
(Kumar A, Crit Care Med 2010)
Un bref passage aux urgences...
Un bref passage aux urgences...
Limitation actuelle: la culture...
Culture
Détection moléculaire directe ?
Détection moléculaire directe ?
1) Préparation
Détection moléculaire directe ?
1) Préparation
2) Amplification/Détection
PCR
Séquençage
ESI-MS
Microarrays
L’étape limitante est….
la préparation de l’échantillon
Avoir au plus vite l’ADN de la bactérie!
Autres méthodes?
PlexID / IRIDICA (Abbott)
Depuis 2009 au GRL (HUG)
Autres méthodes?
Séquenceur de 3ème génération
(Oford Nanopore Technology)
Depuis 2014 au GRL
PlexID / IRIDICA (Abbott)
Depuis 2009 au GRL (HUG)
Sample preparation is the key!
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”
Sample Preparation Work–Flow
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”
MagPhase
Technology
Sample Preparation Work–Flow
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
Next generation Magnetic Particles
Handling Technology
Microfluidic Automation Process
Excellent linearity and reproducibility
DNA extraction from MRSA lysate and detection of
the mecA gene using real-time PCR
Concentration and reproducibility from
large sample volumes
0
20
40
60
80
100
120
140
1 CFU/ml 0.5 CFU/ml 0.3 CFU/ml
E. Coli DNA capture (10 ml sample)
Yie
ldin
%
Sample Preparation Work–Flow
Sample collection Sample Lysis/
homogenization
Target purification
& “preparation”
FibroTrap
Technology
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
FibroTrap technology: Prologue !
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.
Injured site
Microbe
Cas
ca
de
Re
ac
tion
s
Activated coagulation enzyme:
(Serpin -2)
Coagulogen
monomersCoagulin Polymer
FibroTrap technology: Prologue !
Coagulation for
Bacterial Defense
Coagulation for injury
protection
-540 M
years
Injured site
Ca
sc
ad
e
Rea
ctio
ns
Activated coagulation enzyme:
Thrombin
Fibrinogen
monomers
Fibrin Polymer
Pelleting sample preparation technologyA 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
Pelleting sample preparation technologyVacutainer-like technology: potential
combination with various identification methods
Cultures (agar, broth, …)
Micro-cultures
MDx (PCR, sequencing …)
Spectrometry (MALDI, ESI, Raman…)
Sydney Brenner(2002 Nobel prize for Medicine)
Sydney Brenner(2002 Nobel prize for Medicine)
In biotechnology,
the one -"omics"
that really counts is:
economics
Sydney Brenner(2002 Nobel prize for Medicine)
In biotechnology,
the one -"omics"
that really counts is:
economics
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
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