RFP-based method for real-time tracking of invasive bacteria in a heterogeneous population of cells

Akinsola, Rasaq; Adewoyin, Malik; Lee, Choon Weng; Sim, Edmund Ui Hang; Narayanan, K.

doi:10.1016/j.ab.2021.114432

Cited by 1 publication

(2 citation statements)

References 31 publications

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“…Some high-throughput assays have been developed to quantify the extent of bacterial adhesion on host cells. For example, flow cytometry-based adherence assays that quantify interactions between bacteria and eukaryotic cells have been designed for fast and reproducible measurements ( 49 , 66 , 67 ). More recently, fluorescently tagged strains, bacteria carrying fluorescence proteins on plasmids, and live fluorescent stains have been used to quantify the level of adherence with a throughput of 96 samples per assay ( 49 , 66 , 67 ).…”

Section: Adhesion and Invasionmentioning

confidence: 99%

“…For example, flow cytometry-based adherence assays that quantify interactions between bacteria and eukaryotic cells have been designed for fast and reproducible measurements ( 49 , 66 , 67 ). More recently, fluorescently tagged strains, bacteria carrying fluorescence proteins on plasmids, and live fluorescent stains have been used to quantify the level of adherence with a throughput of 96 samples per assay ( 49 , 66 , 67 ). Another method uses virtual colony counts, an absorbance-based measurement showing a good correlation with traditional plate-based colony counts for Salmonella with multiple eukaryotic cell lines ( 30 ).…”

Section: Adhesion and Invasionmentioning

confidence: 99%

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High-throughput functional trait testing for bacterial pathogens

Stromberg,

Phillips,

Omberg

et al. 2023

mSphere

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Functional traits are characteristics that affect the fitness and metabolic function of a microorganism. There is growing interest in using high-throughput methods to characterize bacterial pathogens based on functional virulence traits. Traditional methods that phenotype a single organism for a single virulence trait can be time consuming and labor intensive. Alternatively, machine learning of whole-genome sequences (WGS) has shown some success in predicting virulence. However, relying solely on WGS can miss functional traits, particularly for organisms lacking classical virulence factors. We propose that high-throughput assays for functional virulence trait identification should become a prominent method of characterizing bacterial pathogens on a population scale. This work is critical as we move from compiling lists of bacterial species associated with disease to pathogen-agnostic approaches capable of detecting novel microbes. We discuss six key areas of functional trait testing and how advancing high-throughput methods could provide a greater understanding of pathogens.

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