Selection of Peptidoglycan-Specific Aptamers for Bacterial Cells Identification

被引:29
作者
Ferreira, Ieda Mendes [1 ]
de Souza Lacerda, Camila Maria [1 ]
de Faria, Ligia Santana [1 ]
Correa, Cristiane Rodrigues [1 ]
Ribeiro de Andrade, Antero Silva [1 ]
机构
[1] CDTN, CNEN, BR-31120970 Belo Horizonte, MG, Brazil
关键词
Aptamer; SELEX; Peptidoglycan; Bacterial infection; Radiopharmaceutical; STAPHYLOCOCCUS-AUREUS BACTEREMIA; DNA APTAMERS; LIGANDS; BINDING; SELEX; RADIOPHARMACEUTICALS; ASSAY;
D O I
10.1007/s12010-014-1206-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (K-d) for Antibac1 was 0.415 +/- 0.047 mu M and for Antibac2 was 1.261 +/- 0.280 mu M. These aptamers labeled with P-32 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.
引用
收藏
页码:2548 / 2556
页数:9
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