Leader: Luca Francioso (CNR-IMM); Other collaborator(s): Pietro Aleardo Siciliano (CNR)
This task is devoted to the development of MEMS technologies for implementation of ultra-miniturized biological, chemical and physical sensor array able to guarantee the assessment or monitoring of a safety environment or biological parameters thanks to the high degree of sensors integration; this paradigm can be enabled by micro- and nanotechnologies available at CNR microfabrication labs. The proposed sensors can be integrated with interoperable networks designed in Task 4.2 and benefit from artificial intelligence schemes of task 4.4.
Brief description of the activities and of the intermediate results:
During the report period, the work has been focused on a dedicated optimization of the wearable sensors identified in the previous period, focusing on the fabrication/integration issues in order to guarantee a detection level compatible with target analytes concentrations in the sweat (in case of skin-based sensors) and in the domestic or working environment. About the proposed technology for indoor safety approach, a paper-based technology for an airborne pathogen-monitoring system capable of on-site detection and identification of pathogen was investigated. The technology integrates a sampling/monitoring platform for on-site and real-time detection of airborne viruses an pathogens. The activity has been focused on a paper-based lateral flow assay (LFA) with integrated bioaerosol sampling for the on-site detection and a LAMP-based or PCR-based amplification of genetic proteins in order to dramatically increase the sensitivity of the system.
Main policy, industrial and scientific implications:
Air quality in buildings is a critical determinant of healthy living, especially for populations that are more vulnerable due to their health status or age. Airborne human pathogens and other biological pollutants can impact human health, especially in immunosuppressed persons and older adults, causing infections and respiratory disease. Indoor air quality (IAQ) monitoring, generally, refers to selected chemical pollutants and rarely considers biological contaminants. Having a system that can detect human pathogens in the indoor environment without the need for laboratory analysis and in an autonomous way is an important tool in ensuring a more controlled and safer environment.
Please see the next reporting period.
Brief description of the activities and of the intermediate results:
The paper-based device for airborne pathogen monitoring collects airborne pathogens on a selected paper, a lateral flow module where the sample is lysed, RNA is isolated and then amplified by LAMP on paper. The activities of the reported period focused on the study of the airflow rate for sampling virus particles in an artificial chamber. An artificial air sampler chamber was designed to simulate indoor transmission of pathogens under controllable conditions of pressure, temperature, humidity and airflow. Different papers were selected for the different platform modules based on material and paper properties. The following activities will be related to the sample lysis module, automation of sample transfer and the amplification of nucleic acids on paper. About the cutting-edge equipment implementation, the CNR-IMM will acquire an X-ray diffractometer for chemical characterization of different paper functionalizations and sensors nanomaterials, together with a UV laser and a soft lithography kit for later flow devices microfluidics and rapid prototyping. About the availability of gold standard techniques for biological detection, a qPCR and a Western blotting tools were already purchased by IMM unit.
Brief description of the activities and of the intermediate results:
During the reporting period, work on this task focused on nucleic acid amplification and real-time signal detection on paper. To simulate the detection of indoor airborne pathogens, saliva samples were used for RNA extraction and isolation. Initially, Loop-Mediated Isothermal Amplification (LAMP) assays targeting actin RNA were conducted using a commercial LAMP kit in solution. The reaction was carried out using a specific primer mix, a blend of Bst polymerase and reverse transcriptase in an optimized buffer solution, and a fluorescent dye for real-time fluorescence measurement of LAMP. To assess the feasibility of the reaction on paper, the LAMP solution was dropped to a selected glass fiber pad along with the RNA target. The paper was heated to 65°C for 30 minutes, and a miniature module for real-time fluorescence detection was used to confirm successful RNA target amplification.
Scientific Publications