Rapid detection of <i>Pseudomonas aeruginosa</i> using a DNAzyme‐based sensor
Mingcan Qin, Xiaoyi Ma, Shihui Fan, Hangjie Wu, Wanli Yan, Xiaopeng Tian, Jing Lü, Mingsheng Lyu, Shujun Wang
Abstract
Abstract In the present study, a DNAzyme was screened in vitro through the use of a DNA library and crude extracellular mixture (CEM) of Pseudomonas aeruginosa . Following eight rounds of selection, a DNAzyme termed PAE‐1 was obtained, which displayed high rates of cleavage with strong specificity. A fluorescent biosensor was designed for the detection of P. aeruginosa in combination with the DNAzyme. A detection limit as low as 1.2 cfu/ml was observed. Using proteases and filtration, it was determined that the target was a protein with a molecular weight of 10 kDa–50 kDa. The DNAzyme was combined with a polystyrene board to construct a simple indicator plate sensor which produced a color that identified the target within 10 min. The results were reliable when tap water and food samples were tested. The present study provides a novel experimental strategy for the development of sensors based on a DNAzyme to rapidly detect P. aeruginosa in the field.