RNA-based sandwich hybridisation method for detection of lactic acid bacteria in brewery samples

Sanna Huhtamella, Marika Leinonen, Timo Nieminen, Beatrix Fahnert, Liisa Myllykoski, Antje Breitenstein, Peter Neubauer (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

18 Citations (Scopus)

Abstract

Recently we showed the applicability and sensitivity of the RNA-based sandwich hybridisation assay (SHA) for detection of Gram-negative cells in environmental samples [Leskelä, T., Tilsala-Timisjärvi, A., Kusnetsov, J., Neubauer, P., Breitenstein, A., 2005. Sensitive genus-specific detection of Legionella by a 16S rRNA based sandwich hybridization assay. J. Microbiol. Met. 62, 167–179.]. In this study the aim was to test and optimise this method for the detection of Gram-positive cells from brewery yeast slurries that contain up to 109 yeast cells/ml. Eleven new oligonucleotide probes were designed for group-specific detection of different beer-spoiling lactic acid bacteria of the genera Lactobacillus and Pediococcus. Functionality of the designed probes was shown by testing individual and paired probes using in vitro transcribed 16S rRNA and crude cell extracts as samples. Various simple and fast cell disruption methods were evaluated for the efficient disruption of lactobacilli and pediococci. The applicability of the designed oligonucleotide probes and the SHA for detection of brewery contaminants was demonstrated using both artificial and actual yeast slurry samples from brewery fermentation tanks with either fluorimetric readout or an electric biochip analyser.
Original languageEnglish
Pages (from-to)543-553
JournalJournal of Microbiological Methods
Volume68
Issue number3
DOIs
Publication statusPublished - Mar 2007
MoE publication typeA1 Journal article-refereed

Keywords

  • Lactobacillus brevis
  • Electric biochip
  • Contaminant detection
  • Sandwich hybridisation
  • 16S RNA

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