A novel fluorescence biosensor based on double-stranded DNA branch migration-induced HCR and DNAzyme feedback circuit for sensitive detection of Pseudomonas aeruginosa (clean version)

Pseudomonas aeruginosa (P. aeruginosa) is one of the most common bacteria in nosocomial infection. Here, a novel fluorescence biosensor based on double-stranded DNA branch migration-induced hybridization chain reaction (HCR) and DNAzyme feedback circuit was constructed for sensitive detection of P. aeruginosa. The binding of P. aeruginosa with its aptamer on a DNA three-way junction structure initiated the double-stranded DNA branch migration to form two DNA "Y" junction structures. One DNA "Y" junction structure opened the fluorescence-labelled DNA hairpins and triggered the HCR. The other DNA "Y" junction structure formed a double-stranded DNAzyme and cleaved the specific ribonucleotide site, producing new triggering probes to start the next cycle of the double-stranded DNA branch migration. Ultimately, a large number of DNA "Y" junction structures were produced, which greatly promoted signal amplification. Under optimized conditions, the proposed biosensor detected a wide linearity range of 10²-10⁷ CFU mL^-1, and the limit of detection was 37 CFU mL^-1 (S/N = 3). The recovery test results indicated that the biosensor has promising clinical application potential. Because of the simultaneous initiation of the HCR and the DNAzyme feedback circuit through the double-stranded DNA branch migration, the constructed biosensor provided an ideal platform for pathogenic bacteria detection without protein enzymes and complex signal amplification procedures.