Point-of-Care medical device development for high sensitivity multiplexed detection of blood biomarkers for health care management of cardiac patients ; Développement d'un dispositif médical pour la détection simultanée haute sensibilité de biomarqueurs sanguins pour la prise en charge terrain des patients en souffrance cardiaque

Cardiovascular diseases are the leading cause of death in the world and several biomarkers are clinically used to diagnose patients. Cardiac troponin is currently the gold standard for the diagnostic of acute myocardial infarction (AMI). In hospital emergency departments, testing patient blood for cardiac troponin is crucial to rule-in or rule-out patients. Conventional methods to detect cardiac troponin and other biomarkers are usually performed in central medical laboratories with costly equipment and long analysis time. Point-of-Care devices propose on-site and rapid testing, but these devices currently lack sensitivity. We propose an innovative quantification assay for cardiac biomarkers integrated into a portable and automated instrument for rapid and high sensitivity testing.We developed a method based on isothermal nucleic acids amplification (LAMP) using an original structure called dumbbell. For the quantification of biomarkers, the sample is incubated with this dumbbell structure containing various types of molecular probes such as aptamers or antibodies. Complete detection protocols are derived for various types of targets and integrated on a portable and automated platform. This latter contains microfluidic cartridges including a network of fluidic chambers used for the discrete manipulation of fluids. The cartridge architecture is designed specifically for the developed assay based on dumbbell amplification, and allow carrying out all the assay steps, from the sample recovery to the result.We validated the isothermal amplification of the dumbbell structure with various types of molecular probes. We studied different parameters influencing the amplification reaction for a better understanding of the amplification process. We used thrombin as a model protein and included a specific thrombin aptamer in the dumbbell structure to obtain sensitive (100 pM) and antibody-free detection using this aptameroLAMP method. It is however limited to targets for which a double aptamer sandwich is available. For the ...