Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity

Katie S Flenker; Elliot L Burghardt; Nirmal Dutta; William J Burns; Julia M Grover; Elizabeth J Kenkel; Tyler M Weaver; James Mills; Hyeon Kim; Lingyan Huang; Richard Owczarzy; Catherine A Musselman; Mark A Behlke; Bradley Ford; James O McNamara

doi:10.1016/j.ymthe.2017.03.015

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Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity

Journal article

Open access

Peer reviewed

Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity

Katie S Flenker, Elliot L Burghardt, Nirmal Dutta, William J Burns, Julia M Grover, Elizabeth J Kenkel, Tyler M Weaver, James Mills, Hyeon Kim, Lingyan Huang, …

Molecular therapy, Vol.25(6), pp.1353-1362

06/07/2017

DOI: 10.1016/j.ymthe.2017.03.015

PMCID: PMC5474879

PMID: 28391960

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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 . McNamara and colleagues developed a rapid, ultrasensitive assay that detects E. coli by measuring the activity of endonuclease I, an E. coli nuclease. The assay detected clinically significant levels of E. coli in patient urine samples, suggesting its utility as a rapid diagnostic for urinary tract infections.

diagnostics

nuclease

Escherichia coli

endonuclease I

DNase

Original

deoxyribonuclease

urinary tract infection

ribonuclease

RNase

oligonucleotide

Details

Title: Subtitle: Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity
Creators: Katie S Flenker - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Elliot L Burghardt - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Nirmal Dutta - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
William J Burns - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Julia M Grover - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Elizabeth J Kenkel - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Tyler M Weaver - Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
James Mills - Department of Psychiatry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Hyeon Kim - University of Iowa Research Foundation, University of Iowa, Iowa City, IA 52242, USA
Lingyan Huang - Integrated DNA Technologies (IDT), Coralville, IA 52241, USA
Richard Owczarzy - Integrated DNA Technologies (IDT), Coralville, IA 52241, USA
Catherine A Musselman - Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Mark A Behlke - Integrated DNA Technologies (IDT), Coralville, IA 52241, USA
Bradley Ford - Department of Pathology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
James O McNamara - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Resource Type: Journal article
Publication Details: Molecular therapy, Vol.25(6), pp.1353-1362
DOI: 10.1016/j.ymthe.2017.03.015
PMID: 28391960
PMCID: PMC5474879
NLM abbreviation: Mol Ther
ISSN: 1525-0016
eISSN: 1525-0024
Publisher: American Society of Gene & Cell Therapy
Grant note: DOI: 10.13039/100012483, name: University of Georgia Research Foundation; DOI: 10.13039/100000001, name: National Science Foundation, award: 1452411; DOI: 10.13039/100000002, name: National Institutes of Health, award: AI101391, AI106738
Language: English
Date published: 06/07/2017
Academic Unit: Microbiology and Immunology; Pathology; Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9984047740002771

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