Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing

Emanuela Altobelli, Ruchika Mohan, Kathleen E. Mach, Mandy Lai Yi Sin, Joseph C. Liao, Maurizio Buscarini, Pak Kin Wong, Vincent Gau, Niaz Banaei, Joseph C. Liao

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background Standard diagnosis of urinary tract infection (UTI) via urine culture for pathogen identification (ID) and antimicrobial susceptibility testing (AST) takes 2–3 d. This delay results in empiric treatment and contributes to the misuse of antibiotics and the rise of resistant pathogens. A rapid diagnostic test for UTI may improve patient care and antibiotic stewardship. Objective To develop and validate an integrated biosensor assay for UTI diagnosis, including pathogen ID and AST, with determination of the minimum inhibitory concentration (MIC) for ciprofloxacin. Design, setting, and participants Urine samples positive for Enterobacteriaceae (n = 84) or culture-negative (n = 23) were obtained from the Stanford Clinical Microbiology Laboratory between November 2013 and September 2014. Each sample was diluted and cultured for 5 h with and without ciprofloxacin, followed by quantitative detection of bacterial 16S rRNA using a single electrochemical biosensor array functionalized with a panel of complementary DNA probes. Pathogen ID was determined using universal bacterial, Enterobacteriaceae (EB), and pathogen-specific probes. Phenotypic AST with ciprofloxacin MIC was determined using an EB probe to measure 16S rRNA levels as a function of bacterial growth. Measurements Electrochemical signals for pathogen ID at 6 SD over background were considered positive. An MIC signal of 0.4 log units lower than the no-antibiotic control indicated sensitivity. Results were compared to clinical microbiology reports. Results and limitations For pathogen ID, the assay had 98.5% sensitivity, 96.6% specificity, 93.0% positive predictive value, and 99.3% negative predictive value. For ciprofloxacin MIC the categorical and essential agreement was 97.6%. Further automation, testing of additional pathogens and antibiotics, and a full prospective study will be necessary for translation to clinical use. Conclusions The integrated biosensor platform achieved microbiological results including MIC comparable to standard culture in a significantly shorter assay time. Further assay automation will allow clinical translation for rapid molecular diagnosis of UTI. Patient summary We have developed and validated a biosensor test for rapid diagnosis of urinary tract infections. Clinical translation of this device has the potential to significantly expedite and improve treatment of urinary tract infections. We developed an integrated biosensor assay based on electrochemical detection of bacterial 16S rRNA for simultaneous uropathogen identification and phenotypic antimicrobial susceptibility testing. Validation using clinical urine samples demonstrated outstanding diagnostic accuracy and the potential to significantly accelerate diagnosis of urinary tract infection to promote better antibiotic stewardship.

Original languageEnglish (US)
Pages (from-to)293-299
Number of pages7
JournalEuropean Urology Focus
Volume3
Issue number2-3
DOIs
StatePublished - Apr 1 2017

Fingerprint

Biosensing Techniques
Urinary Tract Infections
Microbial Sensitivity Tests
Ciprofloxacin
Anti-Bacterial Agents
Enterobacteriaceae
Automation
Urine
Microbiology
DNA Probes
Routine Diagnostic Tests
Patient Care
Complementary DNA
Prospective Studies
Sensitivity and Specificity
Equipment and Supplies
Therapeutics
Growth

Keywords

  • Biosensing techniques
  • Microbial sensitivity tests
  • Molecular diagnostics
  • Point-of-care system
  • Urinary tract infection

ASJC Scopus subject areas

  • Urology

Cite this

Altobelli, E., Mohan, R., Mach, K. E., Sin, M. L. Y., Liao, J. C., Buscarini, M., ... Liao, J. C. (2017). Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing. European Urology Focus, 3(2-3), 293-299. https://doi.org/10.1016/j.euf.2015.12.010

Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing. / Altobelli, Emanuela; Mohan, Ruchika; Mach, Kathleen E.; Sin, Mandy Lai Yi; Liao, Joseph C.; Buscarini, Maurizio; Wong, Pak Kin; Gau, Vincent; Banaei, Niaz; Liao, Joseph C.

In: European Urology Focus, Vol. 3, No. 2-3, 01.04.2017, p. 293-299.

Research output: Contribution to journalArticle

Altobelli, E, Mohan, R, Mach, KE, Sin, MLY, Liao, JC, Buscarini, M, Wong, PK, Gau, V, Banaei, N & Liao, JC 2017, 'Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing', European Urology Focus, vol. 3, no. 2-3, pp. 293-299. https://doi.org/10.1016/j.euf.2015.12.010
Altobelli, Emanuela ; Mohan, Ruchika ; Mach, Kathleen E. ; Sin, Mandy Lai Yi ; Liao, Joseph C. ; Buscarini, Maurizio ; Wong, Pak Kin ; Gau, Vincent ; Banaei, Niaz ; Liao, Joseph C. / Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing. In: European Urology Focus. 2017 ; Vol. 3, No. 2-3. pp. 293-299.
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AU - Buscarini, Maurizio

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N2 - Background Standard diagnosis of urinary tract infection (UTI) via urine culture for pathogen identification (ID) and antimicrobial susceptibility testing (AST) takes 2–3 d. This delay results in empiric treatment and contributes to the misuse of antibiotics and the rise of resistant pathogens. A rapid diagnostic test for UTI may improve patient care and antibiotic stewardship. Objective To develop and validate an integrated biosensor assay for UTI diagnosis, including pathogen ID and AST, with determination of the minimum inhibitory concentration (MIC) for ciprofloxacin. Design, setting, and participants Urine samples positive for Enterobacteriaceae (n = 84) or culture-negative (n = 23) were obtained from the Stanford Clinical Microbiology Laboratory between November 2013 and September 2014. Each sample was diluted and cultured for 5 h with and without ciprofloxacin, followed by quantitative detection of bacterial 16S rRNA using a single electrochemical biosensor array functionalized with a panel of complementary DNA probes. Pathogen ID was determined using universal bacterial, Enterobacteriaceae (EB), and pathogen-specific probes. Phenotypic AST with ciprofloxacin MIC was determined using an EB probe to measure 16S rRNA levels as a function of bacterial growth. Measurements Electrochemical signals for pathogen ID at 6 SD over background were considered positive. An MIC signal of 0.4 log units lower than the no-antibiotic control indicated sensitivity. Results were compared to clinical microbiology reports. Results and limitations For pathogen ID, the assay had 98.5% sensitivity, 96.6% specificity, 93.0% positive predictive value, and 99.3% negative predictive value. For ciprofloxacin MIC the categorical and essential agreement was 97.6%. Further automation, testing of additional pathogens and antibiotics, and a full prospective study will be necessary for translation to clinical use. Conclusions The integrated biosensor platform achieved microbiological results including MIC comparable to standard culture in a significantly shorter assay time. Further assay automation will allow clinical translation for rapid molecular diagnosis of UTI. Patient summary We have developed and validated a biosensor test for rapid diagnosis of urinary tract infections. Clinical translation of this device has the potential to significantly expedite and improve treatment of urinary tract infections. We developed an integrated biosensor assay based on electrochemical detection of bacterial 16S rRNA for simultaneous uropathogen identification and phenotypic antimicrobial susceptibility testing. Validation using clinical urine samples demonstrated outstanding diagnostic accuracy and the potential to significantly accelerate diagnosis of urinary tract infection to promote better antibiotic stewardship.

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