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Development of a fully automated, integrated solution that can support a wide range of complex sample preparation protocols

Final Report Summary - SPCARD (Development of a fully automated, integrated solution that can support a wide range of complex sample preparation protocols)

Sepsis is defined as a life threatening condition that arises when the body’s response to an infection spirals out of control. It represents a global health problem with a very high mortality ranging from 15% to 50%. Each year the number of cases of severe sepsis is rising. Currently 10-14% of all ICU admissions in the western world are caused by sepsis.
The major unmet clinical need is a fast diagnosis that gives insight in the pathogen identification and antibiotic resistance. Currently culturing is the gold standard. However, this method requires 24-48h to provide these answers. Molecular diagnosis offers the potential to provide these answers in a couple of hours. The missing link to provide a good molecular diagnosis is the capability to enrich the minute amount of pathogens from large volumes of blood and bring the technology to the point of need.
The aim of the fellowship was to optimize and develop a disposable cartridge and instrument to perform automated pathogen DNA enrichment from large volumes of blood. This would enable a fast and accurate diagnosis of the causative pathogen.
The starting point of the fellowship was an early research cartridge and instrument that could perform the sample preparation. During the fellowship, this early stage proof of concept has been transformed into a full functional prototype cartridge and instrument which can be manufactured in larger numbers and be used in the clinical routine workflow. The main improvements were:
For the instrument:
- Integration of electronics and software
- Integration of liquid sensing element to enable measurement of liquid levels
- Integration of intuitive user interaction (LED progress bar, display)
- Integration of pressure/vacuum pump. The system only requires a power supply
For the cartridge:
- Integration of a reagent container to store all reagents during shelf life
- Optimization of the entire assembly and chemicals to withstand gamma irradiation to remove any contaminating DNA background
- Simple interface to plug in a blood tube. There is no need to open the blood tube.
- Sample processing in 30-35 min with only 2 min hands on time from the operator
- Optimization of the assembly and manufacturing methods to allow annual production up to 40.000 pieces per year
All these changes have resulted in a simple instrument that requires only a power connection. An experiment can be performed by minimally trained staff with only 2 minutes hands-on time. The LED progress bar allows easy monitoring of the status of the instrument.
The sample output of this cartridge can be further processed using a standard molecular analysis (DNA extraction and amplification). The results indicated that it is feasible to detect an array of different bacteria and fungi at very low concentrations (1-10 CFU/ml). The cartridge is exposed to gamma irradiation to degrade all pathogenic DNA that might be introduced during the manufacturing, thus reducing the potential for false negatives. Despite exposure to this harsh treatment the cartridge has shown no loss in process efficacy.
During the Marie curie fellowship a business plan aimed at future commercialization has been worked out in detail. This included also a detailed financial plan and prediction of the potential market and market size. It has become clear that any molecular test in the sepsis field will face huge challenges to get widely accepted. This is because the current gold standard relies on culturing and not on a molecular test. However the advantages of a molecular test are well known and recognized throughout the key-opinion leaders (KOLs) in the field. Therefore our strategy in the initial years is based on a close collaboration with a limited number of highly influential KOLs. Therefore Polaris is currently being used in several studies in the Netherlands including the CTMM MARS trial, BLENT and in collaboration with Jeroen Bosh Hospital. In the next year, the number of KOLs will be expanded significantly and KOLs from different locations within EU and US will be included.
These collaborations gave us access to real clinical samples and the possibility to align our results with standard culturing results and also medical data (SOFA, APACHE scores,…).