title: Rapid Detection of Urinary Tract Infections via Bacterial Nuclease Activity.
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.
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Katie S
authors.initials: KS
authors.lastname: Flenker
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Burghardt
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Nirmal
authors.initials: N
authors.lastname: Dutta
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: Burns
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Julia M
authors.initials: JM
authors.lastname: Grover
authors.affiliation: Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Kenkel
authors.affiliation: Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Tyler M
authors.initials: TM
authors.lastname: Weaver
authors.affiliation: Department of Psychiatry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: James
authors.initials: J
authors.lastname: Mills
authors.affiliation: University of Iowa Research Foundation, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Hyeon
authors.initials: H
authors.lastname: Kim
authors.affiliation: Integrated DNA Technologies (IDT), Coralville, IA 52241, USA.
authors.forename: Lingyan
authors.initials: L
authors.lastname: Huang
authors.affiliation: Integrated DNA Technologies (IDT), Coralville, IA 52241, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Owczarzy
authors.affiliation: Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Musselman
authors.affiliation: Integrated DNA Technologies (IDT), Coralville, IA 52241, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Behlke
authors.affiliation: Department of Pathology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Bradley
authors.initials: B
authors.lastname: Ford
authors.affiliation: 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.
authors.forename: James O
authors.initials: JO
authors.lastname: McNamara
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D015415:Biomarkers; D003850:Deoxyribonuclease I / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D008836:Micrococcal Nuclease / Q000378:metabolism; D016017:Odds Ratio; D012372:ROC Curve; D015203:Reproducibility of Results; D013211:Staphylococcus aureus / Q000201:enzymology; D014552:Urinary Tract Infections / Q000175:diagnosis* / Q000382:microbiology* / Q000652:urine
chemical_list: D015415:Biomarkers; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease
keywords: DNase; Escherichia coli; RNase; deoxyribonuclease; diagnostics; endonuclease I; nuclease; oligonucleotide; ribonuclease; urinary tract infection