Pre-analytics for pandemic preparedness
PI(s)/Head responsible for the resource:
Martin Sundqvist
Host organisation(s):
Dept. of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital
Resource description:
This project will focus on the crucial pre-analytical steps for detection of viruses and bacteria with metagenomics in order to deliver a national standard operating procedure. The main focus will be to compare and evaluate matrices for sample transportation, effect of storage and nucleic acid extraction methods. The main outcome measure will be the effect on the quality (quantity and fragmentation of DNA/cDNA, the amount of microbial reads, bias in relative composition and ability to use sequences for further typing) of fast long-read sequencing using Oxford Nanopore technology and bioinformatics tools. The most likely infection route in coming pandemics is the upper respiratory tract and upper respiratory samples will thus be our primary focus. In the autumn of 2022 the project will focus on methodological issues where exact quantification of SARS-CoV-2, Influenza and RSV using ddPCR will be implemented. Further, samples from the upper airways will be spiked with virus and bacteria, stored in different conditions and then analysed using ddPCR, culture and metagenomic approaches. During the winter of 2022/2023, clinical samples will be collected and analysed to verify these findings. The project aims to expand into more sample matrices and the detection of antibiotic resistance genes and direct typing of viruses and bacteria in 2023. The data from these experiments will render new information regarding the susceptibility of different sample matrices to the environmental conditions.
Contact information:
Martin Sundqvist
Dept. of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital
Email: martin.sundqvist@regionorebrolan.se