Integrated Particle Handling Methods for Multiplexed Microbial Identification and Characterization in Sediments and Groundwater
Molecular methods are still relatively ineffectual for monitoring community dynamics during bioremediation, due primarily to the cost, technical difficulty and retrospective nature of the analyses. For nucleic acid analyses to meaningfully contribute to bioremediation efforts they must not only contribute to the fundamental understanding of microbial ecology, but also be formatted in such a manner that infield analysis can be achieved. The objective of this project is therefore to develop an integrated microbial and nucleic acid detection method and prototype system for the characterization and analysis of subsurface sediments, focusing on the molecular detection of metal and sulfate-reducing bacteria and activity in sediments obtained from the Oak Ridge Field Research Center. We are meeting this objective by combining environmental molecular microbiology with renewable surface techniques, microfluidic systems and microparticle analytical chemistry. The fluidic systems are used to evaluate hypotheses on the integrated biochemistry that is necessary to directly detect 16S rRNA from metal-reducing microbial communities on a suspension microarray, without using the polymerase chain reaction (PCR). These investigations include the use of peptide nucleic acid capture and detection probes and 'tunable surface' concepts to increase nucleic acid capture and detection efficiency and/or mitigate interferences due to co-extracted humic acids. The unified microparticle sample preparation method and suspension array is then used to characterize the 16S rRNA metal- and sulfate-reducing microbial community in FRC sediments before and after biostimulation.
- Research Organization:
- Argonne National Lab. (ANL), Argonne, IL (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Stanford University, Stanford, CA
- Sponsoring Organization:
- USDOE Office of Science (SC)
- OSTI ID:
- 895314
- Report Number(s):
- CONF-NABIR2004-22; TRN: US200702%%749
- Resource Relation:
- Conference: Annual NABIR PI Meeting, March 15-17, 2004, Warrenton, VA
- Country of Publication:
- United States
- Language:
- English
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