dc.contributor.author	Nejati, Fatemeh
dc.contributor.author	Junne, Stefan
dc.contributor.author	Kurreck, Jens
dc.contributor.author	Neubauer, Peter
dc.date.accessioned	2020-08-28T13:10:51Z
dc.date.available	2020-08-28T13:10:51Z
dc.date.issued	2020-06-16
dc.date.updated	2020-06-18T22:57:02Z
dc.description.abstract	Kefir grains are complex microbial systems of several groups of microorganisms. The identification and quantification of the microbial composition of milk kefirs was described in several studies, which provided an insight into the microbial consortia in this complex ecosystem. Nevertheless, the current methods for identification and quantification are not appropriate for deeper studies on kefir consortia, e.g., population dynamics and microbial interactions in kefir grains. This requires another sensitive and reliable quantitative method. Therefore, this study aims to develop multiplexed qPCR assays to specifically detect and quantify, as an example, several microorganisms of the milk kefir microbial community. Primer-probe sets, which target species-specific genes in six bacteria and five yeasts, were designed, and their sensitivity and specificity to the target species was analyzed in simplex as well as four multiplex qPCR assays. The self-designed multiplex assays were applied for the detection of target bacteria and yeast species in milk kefirs, in both, grain and beverage fractions. Detection of all target microorganisms in simplex and multiplex qPCR was achieved by good linearity, efficiency, repeatability and reproducibility in all assays. When the designed assays were applied on six kefirs, all target microorganisms were detected in different samples, but not all in one kefir sample. The two ubiquitous lactobacilli Lactobacillus kefiranofaciens and Lb. kefiri were present in all six kefirs studied, but were associated with different other yeasts and bacteria. Especially on the yeast community a significant diversity was observed. In general, multiplex TaqMan qPCR as developed here was proven to have high potential for specific identification of target microorganisms in kefir samples and for the first time, eleven target bacteria and yeasts of kefir microbiota were rapidly detected and quantified. This study, thus, provides a fast and reliable protocol for future studies on kefir and other similar microbial ecosystems.	en
dc.description.sponsorship	DFG, 414044773, Open Access Publizieren 2019 - 2020 / Technische Universität Berlin	en
dc.identifier.eissn	1664-302X
dc.identifier.uri	https://depositonce.tu-berlin.de/handle/11303/11616
dc.identifier.uri	http://dx.doi.org/10.14279/depositonce-10503
dc.language.iso	en	en
dc.rights.uri	https://creativecommons.org/licenses/by/4.0/	en
dc.subject.ddc	570 Biowissenschaften; Biologie	de
dc.subject.other	TaqMan qPCR	en
dc.subject.other	milk kefir	en
dc.subject.other	quantification	en
dc.subject.other	microbial community	en
dc.subject.other	lactic acid bacteria	en
dc.subject.other	yeast	en
dc.subject.other	Acetobacter	en
dc.title	Quantification of Major Bacteria and Yeast Species in Kefir Consortia by Multiplex TaqMan qPCR	en
dc.type	Article	en
dc.type.version	publishedVersion	en
dcterms.bibliographicCitation.articlenumber	1291	en
dcterms.bibliographicCitation.doi	10.3389/fmicb.2020.01291	en
dcterms.bibliographicCitation.journaltitle	Frontiers in Microbiology	en
dcterms.bibliographicCitation.originalpublishername	Frontiers Media S.A.	en
dcterms.bibliographicCitation.originalpublisherplace	Lausanne	en
dcterms.bibliographicCitation.volume	11	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

Quantification of Major Bacteria and Yeast Species in Kefir Consortia by Multiplex TaqMan qPCR

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