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Projeto n.º 029 - Apoio Especial RESEARCH 4 COVID 19

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Código da Operação 029_Research4Covid-19

Título: Implementation of rapid endpoint PCR systems for SARS-CoV-2 detection in conventional molecular biology laboratories

Síntese do Projeto: We have recently built a free database (CoV2ID) for evaluation of SARS CoV-2 detection protocols (http://covid.portugene.com/), as we have done previously for Ebola and HIV (see coordinator's CV for all publications and links). The database evaluates WHO diagnostic protocols according to the virus genetic diversity, allowing the detection of mutations that may lead to false negative results. We will now include a Blog and Feedback sections for laboratories share their experience and doubts. We will also constantly update the Protocols section with recent improvements on procedures and peer-reviewed evaluations (TASK 1).

Our team has a vast experience in developing molecular kits for biological identifications in different taxonomic groups, including for medical diagnosis and forensic investigations. We have already designed three alternative endpoint PCR protocols for SARS-CoV-2 detection to be tested following WHO recommendations (details can be found here:

https://www.who.int/emergencies/diseases/novel-coronavirus-2019/technical-guidance/laboratory-guidance). We will now acquire and prepare the molecular biology reagents for the endpoint PCR development, including SARS-CoV-2 RNA controls, RNA isolation and reverse transcription kits, etc. (TASK 2). 

The PCRs will be optimized using the synthetic SARS-CoV-2 controls (TASK 3). The protocol will be tested under a variety of conditions to determine its limitations, including for different PCR parameters, direct PCR with no RNA isolation process, different template concentrations, chemical treatments of samples, reproducibility, etc.

After the optimization process, we will test the protocols using real case samples in different laboratories from our partner institutions (TASK 4). We will send reagents and controls and provide the protocol details via our CoV2ID platform. The results will be compared among laboratories. If necessary, adjustments will be made to the protocol following the users'

feedback.

Finally, we will disseminate the details of the most accurate protocols to all national research and clinical laboratories and provide constant feedback on the implementation of protocols, data analyses, etc. We will also organize online or, if possible, in person sessions to help laboratories implement the protocols.

This project is expected to increase the national testing capability allowing the isolation of more infected persons and the understanding of the pandemic progression.

Investigador Responsável: Doutor Filipe Adão Macedo Pereira

Programa de Financiamento: Apoio Especial RESEARCH 4 COVID 19

Instituição Financiadora: FCT - Fundação para a Ciência e a Tecnologia

Data de início: 18-05-2020

Data de conclusão: 31-10-2020

Instituições participantes no Projeto: UC (Proponente);

Custo total elegível (EUR): 17.500,00€

Apoio financeiro público nacional: 17.500,00€

Técnico do Projeto: Daniel Glória

Contacto: 239247021