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A Simple DNAzyme‐Based Fluorescent Assay for Klebsiella pneumoniae
Author(s) -
Ali M. Monsur,
Slepenkin Anatoly,
Peterson Ellena,
Zhao Weian
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800701
Subject(s) - deoxyribozyme , klebsiella pneumoniae , fluorescence , microbiology and biotechnology , bacteria , pathogenic bacteria , chemistry , detection limit , dna , biology , escherichia coli , biochemistry , gene , chromatography , genetics , physics , quantum mechanics
Abstract Pathogenic bacteria pose a serious threat to public health, and the rapid and cost‐effective detection of such bacteria remains a major challenge. Herein, we present a DNAzyme‐based fluorescent paper sensor for Klebsiella pneumoniae . The DNAzyme was generated by an in vitro selection technique to cleave a fluorogenic DNA–RNA chimeric substrate in the presence of K . pneumoniae . The DNAzyme was printed on a paper substrate in a 96‐well format to serve as mix‐and‐read fluorescent assay that exhibits a limit of detection (LOD) 10 5 CFUs mL −1 . Evaluated with 20 strains of clinical bacterial isolates, the DNAzyme produced the desired fluorescence signal with the samples of K . pneumoniae, regardless of their source or drug resistance. The assay is simple to use, rapid, inexpensive, and avoids the complex procedures of sample preparation and equipment. We believe that this DNAzyme‐based fluorescent assay has potential for practical applications to identify K . pneumoniae .