DENVPOC · qPCR Microfluidics point-of-care platform for dengue diagnosis

qPCR Microfluidics point-of-care platform for dengue diagnosis

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.

Data: CORDIS, © European Union