We have developed a convenient and selective method for the detection of Gram-positive bacteria using a ditopic poly(amidoamine) (PAMAM) dendrimer probe. The dendrimer that was modified with dipicolylamine (dpa) and phenylboronic acid groups showed selectivity toward Staphylococcus aureus. The ditopic dendrimer system had higher sensitivity and better pH tolerance than the monotopic PAMAM dendrimer probe. We also investigated the mechanisms of various ditopic PAMAM dendrimer probes and found that the selectivity toward Gram-positive bacteria was dependent on a variety of interactions. Supramolecular interactions, such as electrostatic interaction and hydrophobic interaction, per se, did not contribute to the bacterial recognition ability, nor did they improve the selectivity of the ditopic dendrimer system. In contrast, the ditopic PAMAM dendrimer probe that had a phosphate-sensing dpa group and formed a chelate with metal ions showed improved selectivity toward S. aureus. The results suggested that the targeted ditopic PAMAM dendrimer probe showed selectivity toward Gram-positive bacteria. This study is expected to contribute to the elucidation of the interaction between synthetic molecules and bacterial surface. Moreover, our novel method showed potential for the rapid and species-specific recognition of various bacteria.

The need for a selective bacterial recognition method is evident to overcome the global problem of antibiotic resistance. Even though researchers have focused on boronic acid-based nanoprobes that immediately form boronate esters with saccharides at room temperature, the mechanism has not been well studied. We have developed boronic acid-modified poly(amidoamine) (PAMAM) dendrimers with various surface properties to investigate the mechanism of bacterial recognition. The boronic acid-based nanoprobes showed selectivity toward strains, species, or a certain group of bacteria by controlling their surface properties. Our nanoprobes showed selectivity toward Gram-positive bacteria or Escherichia coli K12W3110 without having to modify the boronic acid recognition sites. The results were obtained in 20 min and visible to the naked eye. Selectivity toward Gram-positive bacteria was realized through electrostatic interaction between the bacterial surface and the positively charged nanoprobes. In this case, the recognition target was lipoteichoic acid on the bacterial surface. On the other hand, pseudo-zwitterionic nanoprobes showed selectivity for E. coli K12W3110, indicating that phenylboronic acid did not recognize the outermost O-antigen on the lipopolysaccharide layer. Boronic acid-based nanoprobes with optimized surface properties are expected to be a powerful clinical tool to recognize multidrug-resistant strains or highly pathogenic bacteria.