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Title: Virulence gene-associated mutant bacterial colonies generate differentiating two-dimensional laser scatter fingerprints
Authors: Atul K. Singh
Lena Leprun
Rishi Drolia
Xingjian Bai
Huisung Kim
Amornrat Aroonnual
Euiwon Bae
Krishna K. Mishra
Arun K. Bhunia
Purdue University
Ivy Tech Community College
CROUS de Dijon
Mahidol University
Keywords: Agricultural and Biological Sciences;Biochemistry, Genetics and Molecular Biology;Environmental Science;Immunology and Microbiology
Issue Date: 1-Jun-2016
Citation: Applied and Environmental Microbiology. Vol.82, No.11 (2016), 3256-3268
Abstract: © 2016, American Society for Microbiology. All Rights Reserved. In this study, we investigated whether a laser scatterometer designated BARDOT (bacterial rapid detection using optical scattering technology) could be used to directly screen colonies of Listeria monocytogenes, a model pathogen, with mutations in several known virulence genes, including the genes encoding Listeria adhesion protein (LAP; lap mutant), internalin A (ΔinlA strain), and an accessory secretory protein (ΔsecA2 strain). Here we show that the scatter patterns of lap mutant, ΔinlA, and ΔsecA2 colonies were markedly different from that of the wild type (WT), with > 95% positive predictive values (PPVs), whereas for the complemented mutant strains, scatter patterns were restored to that of the WT. The scatter image library successfully distinguished the lap mutant and ΔinlA mutant strains from the WT in mixed-culture experiments, including a coinfection study using the Caco-2 cell line. Among the biophysical parameters examined, the colony height and optical density did not reveal any discernible differences between the mutant and WT strains. We also found that differential LAP expression in L. monocytogenes serotype 4b strains also affected the scatter patterns of the colonies. The results from this study suggest that BARDOT can be used to screen and enumerate mutant strains separately from the WT based on differential colony scatter patterns.
ISSN: 10985336
Appears in Collections:Scopus 2016-2017

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