Periodic Reporting for period 2 - MMEM (Miniaturized Matrixed Enthalpy Metter - Portable, affordable, easy to operate, diagnostic device enabling a rapid & accurate diagnosis of a patient infected by antibiotic-resistant bacteria)
Periodo di rendicontazione: 2020-07-01 al 2021-06-30
The European Food and Safety Authority and the ECDC report that antibiotic-resistant bacteria cause 33,000 death/year in the EU alone. This alarming number continuously grows and is forecasted to reach 10 million death/year by 2050. The main threat arises from Carbapenemase Producing Enterobacteriaceae (CPE), which have developed resistance to all kinds of known antibiotics. Infections occur across the continent, and no effective antibiotics are within reach. GenoSmart developing a novel bio-electronic molecular detection system, for detecting the presence of pathogenic DNA obtained from patients. In contrast to all currently available molecular detection techniques, GenoSmart MMEM device aims to fulfill the critical need for rapid, accurate, self-contained diagnosis tool, providing cost-effective test, with no need for any external facility. The sample-to-result diagnostic process will be seamless with no human contact, and will yield clear and simple results in less than 30 minutes.
GenoSmart's ultimate goal planned for this project was to render the MMEM ready for market launch in Europe. This entails a number of technical and strategic objectives, including the completion of the product’s CPE detection capability, the clinical validation through a European clinical study, and exploitation of the results.
GenoSmart's ultimate goal planned for this project was to render the MMEM ready for market launch in Europe. This entails a number of technical and strategic objectives, including the completion of the product’s CPE detection capability, the clinical validation through a European clinical study, and exploitation of the results.
The goal of this project was to develop the MMEM a sample-to-result diagnostic device. The MMEM is comprised of two independent seamlessly connected units. The first unit, termed Automatic Nucleic Acid Extractor (ANAE), extracts genomic DNA from raw bacterial samples and process it into short, clean, CPE specific oligos ready for ensuing molecular reaction. The second unit is GenoSmart’s proprietary sensor, the Miniature Matrixed Enthalpy Meter (MMEM).
During the first year of this project, we developed the ANAE into an automatic machine that isolate DNA from raw samples. The ANAE is comprised from a robotic unit and a perishable cassette. The process begins by inserting a sample into the cassette, the cartridge is introduced into the robotic unit and the DNA is processed into short specific oligos automatically and rapidly with no human contact. We have also developed the Enthalpy Meter to integrate with the sample-to-result device and increased its sensitivity.
During the second year of the project, we further evolved both units, the ANAE and the MEM, and integrated them into one fully automated, seamless, functional sample-to-result device. We have extended the capabilities of the ANAE into a standalone, universal protocol controller, able to run hundreds of independent molecular and biochemical lab procedures in a single, thermocycler-sized machine. The programable ANAE automatically performs the sequential independent steps required for various molecular/biochemical procedures, including liquid transfer and filtering, as well as magnetic separation, sonication, heating, incubation and more, as part of a preloaded or personalized run program. Specifically for this project, the output of the ANAE unit streams a few microliters of sample processed, purified, specific oligos into the MEM unit for detection of the CPE gene in question.
The MEM was modified to integrate into the device, and upon receiving the sample from the ANAE, to start the detection process. Unfortunately, the mechanical, electronical and AI steps taken for improving the sensitivity of the MEM did not meet the required diagnostic sensitivity for detecting CPE in a diagnostic environment.
In addition, we performed a comprehensive bioinformatics survey for optimizing relevant oligonucleotides sequences for detecting the CPE genes most prevalent in Europe. We have also performed regulatory and validation steps for receiving CE mark for the device.
During the first year of this project, we developed the ANAE into an automatic machine that isolate DNA from raw samples. The ANAE is comprised from a robotic unit and a perishable cassette. The process begins by inserting a sample into the cassette, the cartridge is introduced into the robotic unit and the DNA is processed into short specific oligos automatically and rapidly with no human contact. We have also developed the Enthalpy Meter to integrate with the sample-to-result device and increased its sensitivity.
During the second year of the project, we further evolved both units, the ANAE and the MEM, and integrated them into one fully automated, seamless, functional sample-to-result device. We have extended the capabilities of the ANAE into a standalone, universal protocol controller, able to run hundreds of independent molecular and biochemical lab procedures in a single, thermocycler-sized machine. The programable ANAE automatically performs the sequential independent steps required for various molecular/biochemical procedures, including liquid transfer and filtering, as well as magnetic separation, sonication, heating, incubation and more, as part of a preloaded or personalized run program. Specifically for this project, the output of the ANAE unit streams a few microliters of sample processed, purified, specific oligos into the MEM unit for detection of the CPE gene in question.
The MEM was modified to integrate into the device, and upon receiving the sample from the ANAE, to start the detection process. Unfortunately, the mechanical, electronical and AI steps taken for improving the sensitivity of the MEM did not meet the required diagnostic sensitivity for detecting CPE in a diagnostic environment.
In addition, we performed a comprehensive bioinformatics survey for optimizing relevant oligonucleotides sequences for detecting the CPE genes most prevalent in Europe. We have also performed regulatory and validation steps for receiving CE mark for the device.
During the second year of the project, we found that the diagnostic part of our sensor did not achieve the required diagnostic sensitivity. The Automatic DNA Nucleic Acid Extraction (ANAE) unit, on the other hand, was fully developed and proved to isolate short CPE specific DNA oligos from crude bacteria sample within 15 min. We have developed the ANAE into a standalone, universal protocol controller, able to run hundreds of independent molecular and biochemical lab procedures in a single, thermocycler-sized machine. The ANAE automatically performs the sequential independent steps required in various molecular/biochemical procedure, including liquid transfer, filtering, heating, as well as magnetic separation, sonication, heating incubation and many more, as part of a preloaded or personalized run program.
Currently, research and exploratory labs rely on personnel performing repetitive manual protocols involved in molecular and biochemical work with various degrees of skill and results (student, technicians). Next generation lab assays will run differently, and ANAE is a first in kind pioneering system bringing the advantages of all-in-lab, automated, hands-free, 'Lab-on-a-Chip' operation to the researcher's bench. The ANAE will render many of procedures simple, compatible with downstream procedures, adapted to 'Generation Z' expectations and values. The ANAE saves 'hands-on' bench time ensuring traceability of protocols, reduces errors and user-dependency of assay producing repetitive results, protects quality of input and output material and it is green and safe, prevents sample contamination and user exposure.
We expect the ANAE to, for the first time, transform daily lab assays into convenient and automated tasks. From DNA preparation kits to sophisticated CRISPR reactions, cloning and more. Reactions can be run on single or multiple samples as dictated by research needs.
Currently, research and exploratory labs rely on personnel performing repetitive manual protocols involved in molecular and biochemical work with various degrees of skill and results (student, technicians). Next generation lab assays will run differently, and ANAE is a first in kind pioneering system bringing the advantages of all-in-lab, automated, hands-free, 'Lab-on-a-Chip' operation to the researcher's bench. The ANAE will render many of procedures simple, compatible with downstream procedures, adapted to 'Generation Z' expectations and values. The ANAE saves 'hands-on' bench time ensuring traceability of protocols, reduces errors and user-dependency of assay producing repetitive results, protects quality of input and output material and it is green and safe, prevents sample contamination and user exposure.
We expect the ANAE to, for the first time, transform daily lab assays into convenient and automated tasks. From DNA preparation kits to sophisticated CRISPR reactions, cloning and more. Reactions can be run on single or multiple samples as dictated by research needs.