A Simple DNAzyme-Based Fluorescent Assay for Klebsiella pneumoniae

Chembiochem. 2019 Apr 1;20(7):906-910. doi: 10.1002/cbic.201800701. Epub 2019 Feb 11.

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) 105 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.

Keywords: DNAzymes; Klebsiella pneumoniae; fluorescence; paper sensor; pathogenic bacteria.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Typing Techniques / instrumentation
  • Bacterial Typing Techniques / methods*
  • DNA, Catalytic / chemistry*
  • Fluorescent Dyes / chemistry
  • Fluorometry / methods
  • Gene Library
  • Klebsiella pneumoniae / isolation & purification*
  • Limit of Detection
  • Paper

Substances

  • DNA, Catalytic
  • Fluorescent Dyes