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https://www.arca.fiocruz.br/handle/icict/20383
ISOLATION OF AN APTAMER THAT BINDS SPECIFICALLY TO E. COLI.
Affilliation
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil.
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil.
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil./ Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil.
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil.
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil./ Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil
Instituto de Biologia Molecular do Paraná. Department of Research and Development. Curitiba, PR, Brasil.
Abstract
Escherichia coli is a bacterial species found ubiquitously in the intestinal flora of animals,
although pathogenic variants cause major public health problems. Aptamers are short oligonucleotides
that bind to targets with high affinity and specificity, and have great potential for
use in diagnostics and therapy. We used cell-based Systematic Evolution of Ligands by
EXponential enrichment (cell-SELEX) to isolate four single stranded DNA (ssDNA) aptamers
that bind strongly to E. coli cells (ATCC generic strain 25922), with Kd values in the
nanomolar range. Fluorescently labeled aptamers label the surface of E. coli cells, as
viewed by fluorescent microscopy. Specificity tests with twelve different bacterial species
showed that one of the aptamers–called P12-31—is highly specific for E. coli. Importantly,
this aptamer binds to Meningitis/sepsis associated E. coli (MNEC) clinical isolates, and is
the first aptamer described with potential for use in the diagnosis of MNEC-borne
pathologies.
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