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Selection of Peptidoglycan-Specific Aptamers for Bacterial Cells Identification

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Abstract

Peptidoglycan is a highly complex and essential macromolecule of bacterial outer cell wall; it is a heteropolymer made up of linear glycan strands cross-linked by peptides. Peptidoglycan has a particular composition which makes it a possible target for specific bacterial recognition. Aptamers are single-stranded DNA or RNA oligonucleotides that bind to target molecules with high affinity and specificity. Aptamers can be labeled with different radioisotopes and possess several properties that make them suitable for molecular imaging. The purpose of this study was to obtain aptamers for use as radiopharmaceutical in bacterial infection diagnosis. Two aptamers (Antibac1 and Antibac2) against peptidoglycan were selected through the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) methodology. The dissociation constant (Kd) for Antibac1 was 0.415 + 0.047 μM and for Antibac2 was 1.261 + 0.280 μM. These aptamers labeled with 32P showed high affinity for Staphylococcus aureus cells. The binding to S. aureus and Escherichia coli in vitro were significantly higher than for Candida albicans and human fibroblasts, demonstrating their specificity for bacterial cells. These results point Antibac1 and Antibac2 as promising tools for bacterial infections identification.

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Acknowledgments

This research was supported by the Centro de Desenvolvimento da Tecnologia Nuclear/Comissão Nacional de Energia Nuclear (CDTN/CNEN), Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Authors and Affiliations

  1. Centro de Desenvolvimento da Tecnologia Nuclear/Comissão Nacional de Energia Nuclear (CDTN/CNEN), Rua Professor Mário Werneck S/N°, Cidade Universitária-Campus da UFMG, CEP:31120-970, Belo Horizonte, MG, Brasil

    Iêda Mendes Ferreira, Camila Maria de Souza Lacerda, Lígia Santana de Faria, Cristiane Rodrigues Corrêa & Antero Silva Ribeiro de Andrade

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  1. Iêda Mendes Ferreira
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  2. Camila Maria de Souza Lacerda
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  3. Lígia Santana de Faria
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  4. Cristiane Rodrigues Corrêa
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  5. Antero Silva Ribeiro de Andrade
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Correspondence to Antero Silva Ribeiro de Andrade.

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Ferreira, I.M., de Souza Lacerda, C.M., de Faria, L.S. et al. Selection of Peptidoglycan-Specific Aptamers for Bacterial Cells Identification. Appl Biochem Biotechnol 174, 2548–2556 (2014). https://doi.org/10.1007/s12010-014-1206-6

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  • DOI: https://doi.org/10.1007/s12010-014-1206-6

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