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Updated: Aug 7, 2026
Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization
Published on: April 14, 2016
Design and evaluation of 16S rRNA sequence based oligonucleotide probes for the detection and quantification of
Stephan Bathe1, Martina Hausner
1Institute of Water Quality Control and Waste Management, Technical University of Munich, Am Coulombwall, 85748 Garching, Germany. steph-b@gmx.de
Background:
The beta-proteobacterial species Comamonas testosteroni is capable of biotransformation and also biodegradation of a range of chemical compounds and thus potentially useful in chemical manufacturing and bioremediation. The ability to detect and quantify members of this species in mixed microbial communities thus may be desirable.
Results:
We have designed an oligonucleotide probe for use in fluorescent in situ hybridization (FISH) and two pairs of PCR primers targeting a C. testosteroni subgroup. The FISH probe and one of the PCR primer pairs are suitable for quantification of C. testosteroni in mixed microbial communities using FISH followed by quantitative image analysis or real-time quantitative PCR, respectively. This has been shown by analysis of samples from an enrichment of activated sludge on testosterone resulting in an increase in abundance and finally isolation of C. testosteroni. Additionally, we have successfully used quantitative PCR to follow the C. testosteroni abundance during a laboratory scale wastewater bioaugmentation experiment.
Conclusion:
The oligonucleotides presented here provide a useful tool to study C. testosteroni population dynamics in mixed microbial communities.
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