Abstract:
: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.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Paradh AD,Hill AE,Mitchell WJdoi
10.1016/j.mimet.2013.10.014subject
Has Abstractpub_date
2014-01-01 00:00:00pages
25-34eissn
0167-7012issn
1872-8359pii
S0167-7012(13)00329-1journal_volume
96pub_type
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