: Urinary tract infection is the most common urological disease in the United States and is a major cause of patient morbidity and health-care expenditure. This Bioengineering Research Partnership proposal involves development of a microelectromechanical system for the genotypic detection and species-specific identification of uropathogens within a time frame (5-10 minutes from sample collection to readout) that would enable point-of-care diagnosis and treatment. The focus of this proposal is to produce a self-contained microbial pathogen detection device and to examine its performance using clinical urine samples. Research at UCLA has provided two key technological advances that make development of a uropathogen sensor feasible. The first is microfluidics for sample processing. The second is an electrochemical microsensor which allows ultrasensitive detection of specific DNA-RNA or DNA-DNA hybridization events, without the need for target amplification. This project has been in development for over a year involving a multidisciplinary effort including leaders in the fields of microfluidics and microsensor technology, molecular microbiology, pediatrics and biomathematics.
Specific Aim 1 describes how microfluidics studies will be applied to development of a crossflow filter for uropathogen concentration, micromixing for processing of uropathogen nucleic acids, and washing of the sensor surface.
Specific Aim 2 involves fabrication of the microsensor array, development of a streptavidin self-assembled monolayer, and testing of oligonucleotide probes for electrochemical detection of uropathogen rRNA and mRNA on the microsensor surface.
Specific Aim 3 will involve integration of the microfluidics and sensor components and testing of its analytic validity on simulated and actual urine specimens.
Specific Aim 4 will involve batch fabrication of the device and clinical examination of the association between urosensor results and clinical correlates of urinary tract infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB000127-02
Application #
6609681
Study Section
Special Emphasis Panel (ZRG1-SSS-M (02))
Program Officer
Korte, Brenda
Project Start
2002-07-15
Project End
2007-06-30
Budget Start
2003-07-01
Budget End
2004-06-30
Support Year
2
Fiscal Year
2003
Total Cost
$1,279,061
Indirect Cost
Name
University of California Los Angeles
Department
Urology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Lawi, Walson; Wiita, Chris; Snyder, Steven T et al. (2009) A Microfluidic Cartridge System for Multiplexed Clinical Analysis. JALA Charlottesv Va 14:407-412
Mastali, Mitra; Babbitt, Jane T; Li, Yang et al. (2008) Optimal probe length and target location for electrochemical detection of selected uropathogens at ambient temperature. J Clin Microbiol 46:2707-16
Tabibian, James H; Gornbein, Jeffrey; Heidari, Arash et al. (2008) Uropathogens and host characteristics. J Clin Microbiol 46:3980-6
Ivancic, Vesna; Mastali, Mitra; Percy, Neil et al. (2008) Rapid antimicrobial susceptibility determination of uropathogens in clinical urine specimens by use of ATP bioluminescence. J Clin Microbiol 46:1213-9
Gau, Vincent; Wong, David (2007) Oral fluid nanosensor test (OFNASET) with advanced electrochemical-based molecular analysis platform. Ann N Y Acad Sci 1098:401-10
Liao, Joseph C; Mastali, Mitra; Gau, Vincent et al. (2006) Use of electrochemical DNA biosensors for rapid molecular identification of uropathogens in clinical urine specimens. J Clin Microbiol 44:561-70
Gau, Vincent; Ma, Shu-Ching; Wang, Hua et al. (2005) Electrochemical molecular analysis without nucleic acid amplification. Methods 37:73-83
Sun, Chien-Pin; Liao, Joseph C; Zhang, Yao-Hua et al. (2005) Rapid, species-specific detection of uropathogen 16S rDNA and rRNA at ambient temperature by dot-blot hybridization and an electrochemical sensor array. Mol Genet Metab 84:90-9
Suchard, Marc A; Weiss, Robert E; Sinsheimer, Janet S (2005) Models for estimating bayes factors with applications to phylogeny and tests of monophyly. Biometrics 61:665-73