2007 NSTI Nanotechnology Conference and Trade Show - Nanotech 2007 - 10th Annual

An in situ synthesized microfluidic DNA biochip to detect 20 waterborne pathogens

D.M. Tourlousse, A. Munir, R.D. Stedtfeld, F. Ahmad, T. Stedtfeld, A.B. Herzog, J.-M. Rouillard, E. Gulari, J.M. Tiedje and S.A. Hashsham
Michigan State University, US

Keywords:
pathogens, biochip, microfluidic, water, virulence

Abstract:
Parallel and cost-effective detection of pathogens is of interest in many areas including clinical diagnostics, food and water safety, and biodefense. This study reports the development and validation of a microfluidic biochip containing approximately 10,000 in situ synthesized oligonucleotide probes for simultaneous detection of 20 waterborne pathogens. Each pathogen was targeted with 3 to 6 virulence and marker genes. Use of virulence and marker genes as opposed to the 16S rRNA gene used by several previous studies, and use of 5 to 20 oligonucleotide probes per gene are among the key attributes of this biochip. For enhanced sensitivity, multiplex amplification of up to 25 gene targets in a single polymerase chain reaction tube was achieved followed by fluorescent labeling and hybridized to the biochip. Detection limits were estimated to be in the range of 0.01 to 0.001% depending on the GC content of the target gene. Models were developed to assess hybridization time and validated by experimental data. The biochip has the potential to serve as a powerful diagnostic tool for the detection and identification of microorganisms. Considering analysis cost alone, it also provides significant cost savings when samples must be screened for multiple targets ($10-$15 per pathogen).

Back to Program

Sessions Sunday Monday Tuesday Wednesday Thursday Authors Keywords

Nanotech 2007 Conference Program Abstract

 
 

Names, and logos of other organizations are the property of those organizations and not of NSTI.
This event is not open to the general public and NSTI reserves the right to refuse admission and participation to any individual.