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Proceedings Paper

Integrated systems for DNA sample preparation and detection in environmental samples
Author(s): Cynthia J. Bruckner-Lea; Norman C. Anheier; David A. Holman; Toyoko Tsukuda; Mark T. Kingsley; Fred J. Brockman; John M. Price; Jay W. Grate; Darrell P. Chandler
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Paper Abstract

Field-portable sensor system are currently needed for the detection and characterization of biological pathogens in the environment. Nucleic acid analysis is frequently the method of choice for discriminating between pathogenic and non-pathogenic bacteria in environmental samples, however standard protocols are difficult to automate and current microfluidic devices are not configured to analyze environmental samples. In this paper, we describe an automated DNA sample processing system and demonstrate its use for the extraction of bacterial DNA form water and sediment samples. Two challenges in environmental sample analysis are the need to process relatively large sample volumes in order to obtain detectable quantities of DNA present at low concentrations, and the need to purify DNA form a complex sample matrix for downstream detection. These problems are addressed by using sequential injection fluid handling techniques for precise manipulation of the required volumes, and renewable separation columns for automatically trapping and releasing microparticles that are used for sample purification. The renewable microcolumns are used for both bacterial cell concentration and DNA purification. The purified bacterial DNA is then amplified using an on-line PCR module in order to produce detectable quantities of the target DNA.

Paper Details

Date Published: 29 December 2000
PDF: 8 pages
Proc. SPIE 4200, Biochemical and Biomolecular Sensing, (29 December 2000); doi: 10.1117/12.411714
Show Author Affiliations
Cynthia J. Bruckner-Lea, Pacific Northwest National Lab. (United States)
Norman C. Anheier, Pacific Northwest National Lab. (United States)
David A. Holman, Pacific Northwest National Lab. (United States)
Toyoko Tsukuda, Pacific Northwest National Lab. (United States)
Mark T. Kingsley, Pacific Northwest National Lab. (United States)
Fred J. Brockman, Pacific Northwest National Lab. (United States)
John M. Price, Pacific Northwest National Lab. (United States)
Jay W. Grate, Pacific Northwest National Lab. (United States)
Darrell P. Chandler, Pacific Northwest National Lab. (United States)


Published in SPIE Proceedings Vol. 4200:
Biochemical and Biomolecular Sensing
Robert A. Lieberman, Editor(s)

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