Fully automated and integrated Microfluidic Platform for Real-time Molecular Diagnosis of Methicillin-resistant Staphylococcus aureus
Methicillin-resistant Staphylococcus aureus
(MRSA), a virulent organism resistant to
many drugs, is responsible for most nosocomial
and community-acquired infections.
It can cause life-threatening disease, and
treatment options are limited. Effective
diagnostics is a strategic key element in the
campaign against the spread of MRSA,
allowing better infection surveillance and
control measures as well as more efficient
patient treatment and/or isolation options.
We collaborate on the development of a novel process flow enabling prototyping
of microfluidic cartridges made out of polymer films. The wall thicknesses are smaller
than 188 µm, thus enabling efficient thermocycling during real-time polymerase
chain reaction (PCR).
The microfluidic cartridges are designed for operation in a slightly modified
commercial thermocycling instrument. This approach demonstrates new opportunities
for both microfluidic developments and well-established laboratory instruments.
The microfluidic protocol is controlled by centrifugal forces
and divides the liquid sample parallely into independent aliquots of 9.8 µl.
The MagRSA project aims at the development of a
new diagnostics platform that will provide a fast,
simple, automated and accurate identification of
MRSA from clinical samples.
The diagnostic protocol that is proposed relies on
a new and clinically validated procedure that consists
of a direct one-step enrichment of MRSA
present in either nasal or inguinal swabs, followed
by DNA extraction of immunocaptured bacteria
and their identification by multiplex sequence
amplification, using real-time quantitative PCR.
This protocol will be implemented with a simple, hands-off, system based on:
- novel strategies for the integration of full operations
required for the entire nucleic acid analysis
chain in a microfluidic platform; and
- advanced microfluidic magnetic nanoparticles
manipulation technology allowing efficient capture
and extraction of target bacteria and
nucleic acids. The separate steps of sample
preparation, signal amplification by multiplex
PCR, and simultaneous detection of multiple
genes, will be performed as one single step
using a ready-to-use disposable fluidic chip.
In light of the above, this project aims to provide
hospitals and care units with a fast, easy and
automated test for the rapid diagnostic of MRSA.
Moreover, the simplicity of the proposed technology
concept, integrating cost effective and widely
available components, allows for the provision of
low cost systems, a prerequisite condition for the
large adoption of molecular tests by hospitals.
The MagRSA project measurable and quantifiable
objectives can be classified in three categories:
- new molecular diagnostics protocol allowing
efficient and reliable MRSA diagnostics and
- new assay reagents including magnetic nanoparticles
for sample preparation and quantitative
PCR (Q-PCR) related reagents;
- fully automated systems, mainly based on
advanced microfluidics and nanoparticles handling
technologies, for MRSA diagnostics and
The MagRSA project will address the unmet need
for new diagnostics tools for management and
control of antimicrobial resistance in general
and MRSA in particular. Moreover, MagRSA project
will provide a diagnostics platform with potential
applications in molecular diagnostics as the
most growing segment within the global in-vitrodiagnostics
This project, funded by a grant from the European Commission 6th framework programme, gathers 6 entities from Switzerland, Germany, France and Sweden:
- Lausanne, Switzerland,
TATAA Biocenter AB
- Göteborg, Sweden,
Laboratory for MEMS Applications
- Freiburg, Germany,
- Pessac, France,
- Lausanne, Switzerland,
and our lab.