Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-8830
Main Title: Quantitative and sensitive RNA based detection of Bacillus spores
Author(s): Osmekhina, Ekaterina
Shvetsova, Antonina
Ruottinen, Maria
Neubauer, Peter
Type: Article
Language Code: en
Abstract: The fast and reliable detection of bacterial spores is of great importance and still remains a challenge. Here we describe a direct RNA-based diagnostic method for the specific detection of viable bacterial spores which does not depends on an enzymatic amplification step and therefore is directly appropriate for quantification. The procedure includes the following steps: (i) heat activation of spores, (ii) germination and enrichment cultivation, (iii) cell lysis, and (iv) analysis of 16S rRNA in crude cell lysates using a sandwich hybridization assay. The sensitivity of the method is dependent on the cultivation time and the detection limit; it is possible to detect 10 spores per ml when the RNA analysis is performed after 6 h of enrichment cultivation. At spore concentrations above 106 spores per ml the cultivation time can be shortened to 30 min. Total analysis times are in the range of 2–8 h depending on the spore concentration in samples. The developed procedure is optimized at the example of Bacillus subtilis spores but should be applicable to other organisms. The new method can easily be modified for other target RNAs and is suitable for specific detection of spores from known groups of organisms.
URI: https://depositonce.tu-berlin.de/handle/11303/9802
http://dx.doi.org/10.14279/depositonce-8830
Issue Date: 11-Mar-2014
Date Available: 14-Aug-2019
DDC Class: 570 Biowissenschaften; Biologie
Subject(s): spore detection
Bacillus subtilis
RNA hybridization
License: https://creativecommons.org/licenses/by/3.0/
Journal Title: Frontiers in Microbiology
Publisher: Frontiers Media S.A.
Publisher Place: Lausanne
Volume: 5
Article Number: 92
Publisher DOI: 10.3389/fmicb.2014.00092
EISSN: 1664-302X
Appears in Collections:FG Bioverfahrenstechnik » Publications

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