Development of first generation in-situ pathogen detection system (Gen1-IPDS) based on NanoGene assay for near real time E. coli O157

H7 detection

Kristi A. Mitchell, Beelee Chua, Ahjeong Son

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We developed the first generation in-situ pathogen detection system (Gen1-IPDS) based on the NanoGene assay for detecting and quantifying Escherichia coli O157:H7 specific eaeA gene. The NanoGene assay employs the hybridization of target DNA with quantum dot labeled magnetic beads and probe DNAs to detect and quantify the target bacterial gene. The Gen1-IPDS is currently capable of executing four key steps required in the NanoGene assay: sample and reagents introduction, DNA hybridization, magnetic separation of complexes, and sample collection. Operational parameters such as magnet position, hybridization buffer composition, hybridization flow rate, and hybridization temperature were investigated. Using the experimentally determined operational parameters, the target gene was successfully quantified (R2=0.97) over a range of six orders of magnitude (10-12 to 10-6molL-1). The limit of detection (LOD) was determined to be 49×10-15molL-1. The specificity was also demonstrated by the differential discrimination of mismatched target DNAs. The NanoGene assay quantification results via Gen1-IPDS were validated by correlation with its laboratory version (R2=0.97).

Original languageEnglish
Pages (from-to)229-236
Number of pages8
JournalBiosensors and Bioelectronics
Volume54
DOIs
Publication statusPublished - 2014 Apr 15

Fingerprint

Escherichia coli O157
Pathogens
Escherichia coli
Assays
DNA
Genes
Bacterial Genes
Quantum Dots
Magnets
DNA Probes
Magnetic separation
Limit of Detection
Buffers
Semiconductor quantum dots
Temperature
Flow rate
Chemical analysis

Keywords

  • DNA hybridization
  • Gene quantification
  • IPDS
  • Magnetic beads
  • NanoGene assay
  • Quantum dot nanoparticles

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry

Cite this

Development of first generation in-situ pathogen detection system (Gen1-IPDS) based on NanoGene assay for near real time E. coli O157 : H7 detection. / Mitchell, Kristi A.; Chua, Beelee; Son, Ahjeong.

In: Biosensors and Bioelectronics, Vol. 54, 15.04.2014, p. 229-236.

Research output: Contribution to journalArticle

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