Exo-III Enzyme-Assisted Triple Cycle Signal Amplifications for Sensitive and Accurate Identification of Pathogenic Bacteria.

Abstract

Early determination of infectious pathogens is vitally important to select appropriate antibiotics, and to manage nosocomial infection. Herein, we propose a target recognition triggered triple signal amplification-based approach for sensitive pathogenic bacteria detection. In the proposed approach, a double-strand DNA probe (capture probe) that is composed of an aptamer sequence and a primer sequence is designed for specific identification of target bacteria and initiation of following triple signal amplification. After recognition of target bacteria, primer sequence is released from capture probe to bind with the designed H1 probe, forming a blunt terminal in the H1 probe. Exonuclease-III (Exo-III enzyme) specifically recognizes the blunt terminal in H1 probe and degrades the sequence from 3' terminal, resulting a single-strand DNA to induce the following signal amplification. Eventually, the approach exhibits a low detection limit of 36cfu/mL with a broad dynamic range. The high selectivity endows the method a promising prospective for clinical sample analysis.