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Mol Ther. 2017 Jun 7;25(6):1353-1362. doi: 10.1016/j.ymthe.2017.03.015. Epub 2017 Apr 5.

Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity.

Author information

1
Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
2
Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
3
Department of Psychiatry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
4
University of Iowa Research Foundation, University of Iowa, Iowa City, IA 52242, USA.
5
Integrated DNA Technologies (IDT), Coralville, IA 52241, USA.
6
Department of Pathology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
7
Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA. Electronic address: james-mcnamara@uiowa.edu.

Abstract

Rapid and accurate bacterial detection methods are needed for clinical diagnostic, water, and food testing applications. The wide diversity of bacterial nucleases provides a rich source of enzymes that could be exploited as signal amplifying biomarkers to enable rapid, selective detection of bacterial species. With the exception of the use of micrococcal nuclease activity to detect Staphylococcus aureus, rapid methods that detect bacterial pathogens via their nuclease activities have not been developed. Here, we identify endonuclease I as a robust biomarker for E. coli and develop a rapid ultrasensitive assay that detects its activity. Comparison of nuclease activities of wild-type and nuclease-knockout E. coli clones revealed that endonuclease I is the predominant DNase in E. coli lysates. Endonuclease I is detectable by immunoblot and activity assays in uropathogenic E. coli strains. A rapid assay that detects endonuclease I activity in patient urine with an oligonucleotide probe exhibited substantially higher sensitivity for urinary tract infections than that reported for rapid urinalysis methods. The 3 hr turnaround time is much shorter than that of culture-based methods, thereby providing a means for expedited administration of appropriate antimicrobial therapy. We suggest this approach could address various unmet needs for rapid detection of E. coli.

KEYWORDS:

DNase; Escherichia coli; RNase; deoxyribonuclease; diagnostics; endonuclease I; nuclease; oligonucleotide; ribonuclease; urinary tract infection

PMID:
28391960
PMCID:
PMC5474879
DOI:
10.1016/j.ymthe.2017.03.015
[Indexed for MEDLINE]
Free PMC Article

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