dc.contributor.author	Osmekhina, Ekaterina
dc.contributor.author	Shvetsova, Antonina
dc.contributor.author	Ruottinen, Maria
dc.contributor.author	Neubauer, Peter
dc.date.accessioned	2019-08-14T14:28:32Z
dc.date.available	2019-08-14T14:28:32Z
dc.date.issued	2014-03-11
dc.date.updated	2019-08-13T14:57:15Z
dc.description.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.	en
dc.identifier.eissn	1664-302X
dc.identifier.uri	https://depositonce.tu-berlin.de/handle/11303/9802
dc.identifier.uri	http://dx.doi.org/10.14279/depositonce-8830
dc.language.iso	en	en
dc.relation.ispartof	10.14279/depositonce-8959	en
dc.rights.uri	https://creativecommons.org/licenses/by/3.0/	en
dc.subject.ddc	570 Biowissenschaften; Biologie	de
dc.subject.other	spore detection	en
dc.subject.other	Bacillus subtilis	en
dc.subject.other	RNA hybridization	en
dc.title	Quantitative and sensitive RNA based detection of Bacillus spores	en
dc.type	Article	en
dc.type.version	publishedVersion	en
dcterms.bibliographicCitation.articlenumber	92	en
dcterms.bibliographicCitation.doi	10.3389/fmicb.2014.00092	en
dcterms.bibliographicCitation.journaltitle	Frontiers in Microbiology	en
dcterms.bibliographicCitation.originalpublishername	Frontiers Media S.A.	en
dcterms.bibliographicCitation.originalpublisherplace	Lausanne	en
dcterms.bibliographicCitation.volume	5	en
tub.accessrights.dnb	free	en
tub.affiliation	Fak. 3 Prozesswissenschaften::Inst. Biotechnologie::FG Bioverfahrenstechnik	de
tub.affiliation.faculty	Fak. 3 Prozesswissenschaften	de
tub.affiliation.group	FG Bioverfahrenstechnik	de
tub.affiliation.institute	Inst. Biotechnologie	de
tub.publisher.universityorinstitution	Technische Universität Berlin	en

Quantitative and sensitive RNA based detection of Bacillus spores

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