Automated microfluidic platform for on field high sensitivity quantification of cardiac troponin I
Résumé
Cardiovascular diseases are the leading cause of death in the world. Cardiac troponin is the gold standard for the diagnostic of myocardial infarction, and testing patient blood for troponin level is crucial to rule-in or rule-out patients. Portable and self-running devices allowing in vitro diagnostics like Point-Of-Care (POC) devices tend to be an interesting method to test patient blood for troponin, but reducing the time and limit of detection of these tests remain currently key points to be addressed. Instead of classical ELISA immuno-assays, we develop an innovative assay to quantify cardiac biomarkers in blood. This assay is based on a molecular sandwich between the target protein of interest and aptamer or antibody probes, followed by an original isothermal LAMP amplification in order to lower the assay limit of detection. We integrate this innovative aptamero- or immuno-LAMP assay in a microfluidic cartridge. This chip is piloted with an automated instrument and carries out every step of the protocol, thus reducing operator manipulations, sample volume and test time The proof of concept of this new sensing method was achieved by aptamero-LAMP on the thrombin protein (a biomarker for coagulation abnormalities) and by immuno-LAMP for troponin protein. In the last case, we have demonstrated a lower limit of detection of 1pM and a wide dynamic range of four orders of magnitude. Combining this innovative assay with the microfluidic technology will allow us to face the challenges required for cardiac Point-of-Care systems.
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