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Enhanced Detection of Bacteria in Environmental Waters: an RNA-based Approach
Citation:
Santodomingo, J. Enhanced Detection of Bacteria in Environmental Waters: an RNA-based Approach. Presented at University of Cincinnati, Cincinnati, OH, October 03, 2014.
Impact/Purpose:
A presentation to students on the work we do in our laboratory
Description:
Molecular assays (i.e., PCR and qPCR) used in microbial water quality studies often target ribosomal RNA genes (rDNA). However, using DNA as the PCR template does not discriminate between active and dead cells. The use of RNA-based detection methods has recently been proposed as RNA is rather unstable outside of the cell and degrade much faster than DNA in nonactive/dead cells. RNA studies can also take advantage of the higher number of rRNA copies in physiologically active cells. The objective of this seminar is to discuss some of the latest results from our laboratory on the use of RNA-based methods as an alternate approach for the detection of active bacterial members of a microbial community. Specifically, we have compared fecal bacteria detection rates using rRNA- vs rDNA-based qPCR assays, and evaluated the use of rRNA/rDNA ratios to access the metabolic status of different fecal bacterial groups. Overall, our data show that in many cases using RNA approaches increased microbial detection of fecal bacteria in environmental waters while also providing quantitative information on the active bacterial fraction. Limitations and future research needs of the use of molecular for environmental monitoring will also be discussed.
