Application of PNA openers for fluorescence-based detection of bacterial DNA

Peptide nucleic acid (PNA) openers have the unique ability to invade double-stranded DNA with high efficiency and sequence specificity, making it possible to detect short (about 20 bp), single-copy bacterial DNA sequences. PNA openers bind to a target signature site on one strand of bacterial DNA, leaving the other strand open for hybridization with a circularizable oligonucleotide probe. The assembled complex serves as a template for rolling circle amplification. The obtained amplicon is decorated with short, single-stranded DNA probes carrying fluorophores and detected via fluorescence microscopy.