Selection of DNA aptamers specific for live Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic pathogen that causes significant morbidity and mortality in immunocompromised patients, particular cystic fibrosis sufferers, burns victims, diabetics and neonates. It thrives in moist places where it forms biofilms that are exceedingly difficult to eradica...

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Bibliographic Details
Published in:PloS one 2017-09, Vol.12 (9), p.e0185385-e0185385
Main Authors: Soundy, Jennifer, Day, Darren
Format: Article
Language:English
Subjects:
Antibiotics
Aptamers
Aptamers, Nucleotide
Bacteria
Bactericidal activity
Binding
Biofilms
Biology and Life Sciences
Biomaterials
Biomedical materials
Biosensors
Cystic fibrosis
Deoxyribonucleic acid
DNA
DNA, Bacterial
Drug resistance
E coli
Flow Cytometry
Gene expression
Genetic aspects
Gram-negative bacteria
Immunocompromised hosts
Lab-On-A-Chip Devices
Library collections
Ligands
Medicine and Health Sciences
Methods
Microbial mats
Microfluidics
Morbidity
Neonates
Nosocomial infections
Nucleic acid aptamers
Opportunist infection
Pseudomonas aeruginosa
Pseudomonas aeruginosa - genetics
Research and Analysis Methods
Salmonella
SELEX Aptamer Technique
Species Specificity
Surgical implants
Water supply
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Summary:Pseudomonas aeruginosa is an opportunistic pathogen that causes significant morbidity and mortality in immunocompromised patients, particular cystic fibrosis sufferers, burns victims, diabetics and neonates. It thrives in moist places where it forms biofilms that are exceedingly difficult to eradicate on hospital surfaces, in water supplies and implanted biomaterials. Using a live cell SELEX approach we selected DNA aptamers to P. aeruginosa grown as biofilms in microfluidic cells. From a pool of aptamer candidates showing tight binding a stem-loop structure was identified as being important for binding. Enhanced binding and increased specificity was achieved by truncating structures and generating chimeric aptamers from the pool of top candidates. The top candidates have low nanomolar binding constants and high discrimination for P. aeruginosa over other Gram-negative bacteria. The aptamers bind both planktonic grown and biofilm grown cells. They do not have intrinsic bacteriostatic or bactericidal activity, but are ideal candidates for modification for use as aptamer-drug conjugates and in biosensors.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0185385