Periodic Reporting for period 1 - DENVPOC (qPCR Microfluidics point-of-care platform for dengue diagnosis)
Periodo di rendicontazione: 2020-05-18 al 2022-05-17
The objective of this project is to develop an universal virus detection platform system for a health basic unit (HBU) with the purpose of infectious disease diagnosis in vitro (DM DIV). This system will use genetic identification based on FASTGENE qPCR technology and integrated with a pre-treatment process. This detection platform would include among the virus detected: dengue, sarscov-2 and so on. This technology development merge Bforcure microfluidics competency by providing microfluidic prototypes which could generate modules for insertion in others POC diagnosis platform for infectious diseases and be commercialized. The work carried out by the beneficiaries specifically in this project was to improve the development of the pre-treatment platform. This platform is designed, by means of microfluidic technology, to inactivate, purify and concentrate virus. This equipment is of vital importance to the overall functioning of the integrated RT-qPCR device in situations where low concentration virus and complex fluids like plasma blood and saliva are analyzed. The inactivation is the first important step, without inactivation, we cannot extract the ARN and, the virus can still contaminate. Purification is the second important step, without this, the sample is not suitable to be analyzed because the presence of Rnases in complex fluids like saliva, plasma, blood hinders the PCR result. Finally, the last step improves the LOD (low limit of detection). In situations where virus concentration in clinical samples is below the detectable level of the PCR equipment. Concentration in this case will help do improve detection and sensibility. The prototype (FASTprep) conception method is mainly based on the use of incremental successive improvements steps both at manufacturability and technology. Fastprep enables purification and concentration of independent semi-automated samples (1 to 4 maximum) in less than 15 minutes. It is based on 2 to 4 modules operating in parallel mode using filtration and biological chemicals in order to purify and extract the virus.
The major achievements of the project were the following:
RNA/ DNA extraction and purification in less that 2 minutes.
Chemical buffers for RNA/ DNA extraction and purification of RNA/DNA samples with saliva developed with (60 -90)% extraction efficiency interval.
The chemical buffer for lysis is also a Viral transport medium (VTM) which at least 24H conservation.
Decontamination protocol without harmful chemicals for prototype decontamination developed with RNA/DNA (0.06/0.1) % retrieval after decontamination compared to a commercial product with (0/0.09)%.
Biological protocol of prototype validation developed.
Low limit of detection established as 100 RNA copies in 4mL of sample.
4 chambers amplification chip developed for the detection of 4 RNA serotypes.
The major achievements of the project were the following:
RNA/ DNA extraction and purification in less that 2 minutes.
Chemical buffers for RNA/ DNA extraction and purification of RNA/DNA samples with saliva developed with (60 -90)% extraction efficiency interval.
The chemical buffer for lysis is also a Viral transport medium (VTM) which at least 24H conservation.
Decontamination protocol without harmful chemicals for prototype decontamination developed with RNA/DNA (0.06/0.1) % retrieval after decontamination compared to a commercial product with (0/0.09)%.
Biological protocol of prototype validation developed.
Low limit of detection established as 100 RNA copies in 4mL of sample.
4 chambers amplification chip developed for the detection of 4 RNA serotypes.
The work performed from the beginning to the end of this project included successful fabrication of a pre-treatment device which concentrate RNA/DNA, one chemical protocol that purifies RNA/DNA , oneprotocol that decontaminates the pre-treatment device, pre-treatment device protocol that validates and characterizes the prototype and successful fabrication a microfluidic chip and instrument prototype that can concentrate RNA/DNA. The PCR module is launched in the market and the pre-treatment module prototype works sucessfully in the detection of sarscov-2 virus in saliva as well as it can extract other virus. For dissemination, Bforcure shoot a video and publish on Youtube to introduce the work to the public: https://www.youtube.com/watch?v=S2A0jgCCUrY(si apre in una nuova finestra). Also, this year Bforcure has won the innovation trophy at JIB(Journées d’inovation en Biologie), new technology for medical diagnostics in 2021 with Chronos.
The Chronos™ Dx platform plus fastprep are an ultra-fast and easy-to-use PCR test tool that delivers results in less than 15 minutes for PCR and less than 2 minutes for extraction and purification of samples like saliva, plasma and blood. it is a friendly-user tool with large resolution and sensitivity designed for infectious diseases diagnotic. Mobile and connected, the Chronos™ Dx platform can be used directly in “satellite laboratories”, delocalized in relation to the main medical biology laboratories, to carry out unit tests of various contagious pathologies.The advantages of the platform are multiple: due to its the price, its physical characteristics, accuracy and connectivity, this equipment can be used in remote areas. Can be used in many areas:
Medical biology laboratories: Quickly perform analyzes of urgent samples without disrupting organization and flows to maintain efficient throughput;
Delocalized laboratories: Easily deploy a diagnostic platform to perform PCR tests near patients outside the laboratory premises;
Health facilities: Reduce the sending of samples to reduce delays in patient care and respond effectively to emergencies;
Trips & events:Dealing with pandemics by rapidly testing departing or arriving travelers and participants in an event with a large audience.
Medical biology laboratories: Quickly perform analyzes of urgent samples without disrupting organization and flows to maintain efficient throughput;
Delocalized laboratories: Easily deploy a diagnostic platform to perform PCR tests near patients outside the laboratory premises;
Health facilities: Reduce the sending of samples to reduce delays in patient care and respond effectively to emergencies;
Trips & events:Dealing with pandemics by rapidly testing departing or arriving travelers and participants in an event with a large audience.