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Rapid detection of Pseudomonas aeruginosa using a DNAzyme-based sensor

Journal 2021 · IF 2.863 · 9(7): 3873-3884 · 发表于 2021 年 6 月 11 日
作者: Mingcan Qin,Xiaoyi Ma,Shihui Fan,Hangjie Wu,Wanli Yan,Xiaopeng Tian,Jing Lu,Mingsheng Lyu,Shujun Wang
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摘要

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.