AMR Detection by dPCR v1

Overview Microbes resistant to antimicrobial agents are a major worldwide health challenge. Rapid detection of antimicrobial resistant pathogens and surveillance efforts are critical. Information about antimicrobial resistance (AMR) genes is available in databases, and quantitative PCR (qPCR) has been proven suitable for detecting AMR genes (Abram et al. 2019; Galhano et al. 2021; Wu et al. 2022). In this research session, we will use the epMotion 5075 and a custom 3D-printed adapter to leverage the power of automation and the new QIAGEN QIAcuity Digital PCR system to detect and quantify a target antimicrobial resistance gene in metagenomic DNA samples from soils and compost. Take a virtual tour of the QIAcuity that we will be using for our research! Digital PCR (dPCR) allows for absolute quantification of template DNA or RNA molecules based on Poisson statistics. But, what does that mean and how does dPCR even work? Read about the fundamentals of dPCR in these articles and watch this short video about dPCR: Read Digital PCR for beginners Read Fundamentals of Digital PCR Watch Principles of dPCR Explained During our second research session, we will set up the epMotion 5075 to dilute metagenomic DNA samples. We will set up dPCR by combining a commercial PCR master mix (includes: dNTPs, polymerase, buffers, Mg2+,water) with our research specific primers, probes, and diluted DNA template to carefully pipette into a QIAGEN Nanoplate with partitions for use in the QIAcuity. We will run dPCR on AMR genes from a mixed population of soil microbes to investigate whether soils used as our samples contain microbes with AMR genes.