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Updated: May 10, 2026
Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications
Published on: November 5, 2020
Development and performance of a quantitative PCR for the enumeration of Bdellovibrionaceae
M Van Essche1, I Sliepen, G Loozen
1Catholic University Leuven, Department of Periodontology, Kapucijnenvoer 7, 3000 Leuven, Belgium. Catholic University Leuven, Centre for Molecular Diagnostics, Herestraat 49, 3000 Leuven, Belgium. The Weizmann Institute of Science, Biological Chemistry, Israel. The Hebrew University of Jerusalem, Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, PO Box 12, Rehovot 76100, Israel. LabMET, Laboratory of Microbial Ecology & Technology, Ghent University, B-9000 Ghent, Belgium.
Abstract:
Quantification of Bdellovibrio-and-like organisms (BALOs) by microbial culturing has a number of substantial drawbacks. Therefore a quantitative PCR (qPCR) assay was designed for the culture-independent enumeration of the Bdellovibrionaceae. After optimization, the dynamic range of the qPCR assay was assessed, the specificity was evaluated and a comparison with quantitative microbial culturing was made. To evaluate the suitability of the qPCR assay for analysing environmental samples, fresh water samples were investigated by microbial culturing and by the qPCR assay. The results revealed a substantial difference between the two techniques and indicate that most Bdellovibrionaceae cells are left undetected in environmental samples when only current microbial culturing techniques are used. The application of this new technique is therefore likely to confirm the hitherto underestimated sizes and roles of predatory bacterial populations in nature.
