Quantitative and sensitive RNA based detection of Bacillus spores

dc.contributor.authorOsmekhina, Ekaterina
dc.contributor.authorShvetsova, Antonina
dc.contributor.authorRuottinen, Maria
dc.contributor.authorNeubauer, Peter
dc.date.accessioned2019-08-14T14:28:32Z
dc.date.available2019-08-14T14:28:32Z
dc.date.issued2014-03-11
dc.date.updated2019-08-13T14:57:15Z
dc.description.abstractThe 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.eissn1664-302X
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/9802
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-8830
dc.language.isoenen
dc.relation.ispartof10.14279/depositonce-8959en
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/en
dc.subject.ddc570 Biowissenschaften; Biologiede
dc.subject.otherspore detectionen
dc.subject.otherBacillus subtilisen
dc.subject.otherRNA hybridizationen
dc.titleQuantitative and sensitive RNA based detection of Bacillus sporesen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.articlenumber92en
dcterms.bibliographicCitation.doi10.3389/fmicb.2014.00092en
dcterms.bibliographicCitation.journaltitleFrontiers in Microbiologyen
dcterms.bibliographicCitation.originalpublishernameFrontiers Media S.A.en
dcterms.bibliographicCitation.originalpublisherplaceLausanneen
dcterms.bibliographicCitation.volume5en
tub.accessrights.dnbfreeen
tub.affiliationFak. 3 Prozesswissenschaften>Inst. Biotechnologie>FG Bioverfahrenstechnikde
tub.affiliation.facultyFak. 3 Prozesswissenschaftende
tub.affiliation.groupFG Bioverfahrenstechnikde
tub.affiliation.instituteInst. Biotechnologiede
tub.publisher.universityorinstitutionTechnische Universität Berlinen
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