Detection of beer spoilage bacteria Pectinatus and Megasphaera with acridinium ester labelled DNA probes using a hybridisation protection assay.

Autor: Paradh AD; International Centre for Brewing and Distilling, School of Life Sciences, Heriot- Watt University, Edinburgh EH14 4AS, UK., Hill AE; International Centre for Brewing and Distilling, School of Life Sciences, Heriot- Watt University, Edinburgh EH14 4AS, UK., Mitchell WJ; International Centre for Brewing and Distilling, School of Life Sciences, Heriot- Watt University, Edinburgh EH14 4AS, UK. Electronic address: W.J.Mitchell@hw.ac.uk.
Jazyk: angličtina
Zdroj: Journal of microbiological methods [J Microbiol Methods] 2014 Jan; Vol. 96, pp. 25-34. Date of Electronic Publication: 2013 Nov 01.
DOI: 10.1016/j.mimet.2013.10.014
Abstrakt: DNA probes specific for rRNA of selected target species were utilised for the detection of beer spoilage bacteria of the genera Pectinatus and Megasphaera using a hybridisation protection assay (HPA). All the probes were modified during synthesis by addition of an amino linker arm at the 5' end or were internally modified by inserting an amine modified thymidine base. Synthesised probes then were labelled with acridinium ester (AE) and purified using reverse phase HPLC. The internally AE labelled probes were able to detect target RNA within the range of 0.016-0.0032pmol. All the designed probes showed high specificity towards target RNA and could detect bacterial contamination within the range of ca. 5×10(2)1×10(3) CFU using the HPA. The developed assay was also compatible with MRS, NBB and SMMP beer enrichment media, routinely used in brewing laboratories.
(© 2013 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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