RES-KIT: Point of care kit for antimicrobial resistance detection
In Europe, 25,000 deaths per year are due to Antimicrobial Resistant (AMR) bacteria, representing the major health threat for the next years. The problem was originated mainly by antibiotics misuse like prescription without proper diagnosis (both in humans and animals). Therefore, having timely information of the microbial susceptibility to antibiotics in each case of infection would be key part of the solution. Up to date, the diagnosis of AMR infections in animal health is slow, time-consuming, expensive and centralized (samples must be sent to an external lab). As a result, clinicians have to wait for a minimum of 36 hours to obtain the laboratory results and to know which antibiotics are effective to start the treatment.
Alternative rapid diagnostic technologies recently released to the market or under development are either not capable of precise quantification (e.g. Polymerase Chain Reaction), or not amenable for on-site testing (e.g. MALDI-TOF), or too expensive for a generalized use (e.g. Next Generation Sequencing).
In recent years, detection of ribosomal RNA has been proposed as a means to quantify and identify bacteria by their 16S rRNA sequences. These rRNA molecules are in high concentration inside the cells (~10.000 copies), so that they can be detected without the need of PCR amplification.
In a preliminary work, using our new microfluidic technology, we have demonstrated quantification of an oligonucleotide sequence with an ELISA-like sandwich assay performed on the surface of microbeads. Our lab-on-a-chip designs allow implementing a complete assay protocol including washing and blocking steps in an automated fashion with a very compact instrument. The high surface to volume ratio of a microbead assay inside microchannels yields increased sensitivity compared to traditional ELISA. Availability of large number of valves within this new microfluidic technology provides high multiplexing capability.
With this technology we will develop a rapid test kit (RES-KIT) for quantification, identification, and antimicrobial susceptibility test (AST) for the treatment of infections. Cystitis in pets will be the first RES-KIT short-term target. Urine is an easy sample to work with and access to veterinary market is rapid thanks to lack of regulation. 9 million pets in Spain are susceptible of suffering cystitis. In the long term the products will also be certified for human use.
This project has been funded by Fons Europeus de Desenvolupament Regional (FEDER) of the European Union, within the framework of Programa operatiu FEDER of Catalunya 2014-2020 with a total of 95.000 €