Direct and sensitive detection of foodborne pathogens within fresh produce samples using a field-deployable handheld device

David J. You, Kenneth J. Geshell, Jeong-Yeol Yoon

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Direct and sensitive detection of foodborne pathogens from fresh produce samples was accomplished using a handheld lab-on-a-chip device, requiring little to no sample processing and enrichment steps for a near-real-time detection and truly field-deployable device. The detection of Escherichia coli K12 and O157:H7 in iceberg lettuce was achieved utilizing optimized Mie light scatter parameters with a latex particle immunoagglutination assay. The system exhibited good sensitivity, with a limit of detection of 10CFUmL -1 and an assay time of <6min. Minimal pretreatment with no detrimental effects on assay sensitivity and reproducibility was accomplished with a simple and cost-effective KimWipes filter and disposable syringe. Mie simulations were used to determine the optimal parameters (particle size d, wavelength λ, and scatter angle θ) for the assay that maximize light scatter intensity of agglutinated latex microparticles and minimize light scatter intensity of the tissue fragments of iceberg lettuce, which were experimentally validated. This introduces a powerful method for detecting foodborne pathogens in fresh produce and other potential sample matrices. The integration of a multi-channel microfluidic chip allowed for differential detection of the agglutinated particles in the presence of the antigen, revealing a true field-deployable detection system with decreased assay time and improved robustness over comparable benchtop systems. Additionally, two sample preparation methods were evaluated through simulated field studies based on overall sensitivity, protocol complexity, and assay time. Preparation of the plant tissue sample by grinding resulted in a two-fold improvement in scatter intensity over washing, accompanied with a significant increase in assay time: ∼5min (grinding) versus ∼1min (washing). Specificity studies demonstrated binding of E. coli O157:H7 EDL933 to only O157:H7 antibody conjugated particles, with no cross-reactivity to K12. This suggests the adaptability of the system for use with a wide variety of pathogens, and the potential to detect in a variety of biological matrices with little to no sample pretreatment.

Original languageEnglish (US)
Pages (from-to)399-406
Number of pages8
JournalBiosensors and Bioelectronics
Volume28
Issue number1
DOIs
StatePublished - Oct 15 2011

Fingerprint

Pathogens
Assays
Equipment and Supplies
Ice Cover
Lettuce
Escherichia coli O157
Light
Latex
Latexes
Washing
Plant Preparations
Escherichia coli
Escherichia coli K12
Microfluidics
Syringes
Tissue
Microspheres
Particle Size
Lab-on-a-chip
Limit of Detection

Keywords

  • Escherichia coli K12
  • Escherichia coli O157:H7
  • Iceberg lettuce
  • Lab on a chip
  • Latex immunoagglutination assay
  • Microfluidic device
  • Mie light scattering

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry

Cite this

Direct and sensitive detection of foodborne pathogens within fresh produce samples using a field-deployable handheld device. / You, David J.; Geshell, Kenneth J.; Yoon, Jeong-Yeol.

In: Biosensors and Bioelectronics, Vol. 28, No. 1, 15.10.2011, p. 399-406.

Research output: Contribution to journalArticle

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