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Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products

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dc.title Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products en
dc.contributor.author Wunderlichová, Leona
dc.contributor.author Buňková, Leona
dc.contributor.author Koutný, Marek
dc.contributor.author Valenta, Tomáš
dc.contributor.author Buňka, František
dc.relation.ispartof Food Microbiology
dc.identifier.issn 0740-0020 OCLC, Ulrich, Sherpa/RoMEO, JCR
dc.date.issued 2013
utb.relation.volume 34
utb.relation.issue 2
dc.citation.spage 268
dc.citation.epage 276
dc.type article
dc.language.iso en
dc.publisher Academic Press en
dc.identifier.doi 10.1016/j.fm.2012.09.024
dc.relation.uri http://ac.els-cdn.com/S0740002012002341/1-s2.0-S0740002012002341-main.pdf?_tid=906ed560-acad-11e2-b4d2-00000aacb35e&acdnat=1366787403_27598ec11c61607000a2e1f0584334af
dc.subject Agmatine en
dc.subject Gram-negative bacteria en
dc.subject Multiplex PCR en
dc.subject PCR en
dc.subject Primers en
dc.subject Putrescine en
dc.description.abstract Formation of biogenic amines may occur in food due to metabolic activities of contaminating Gram-negative bacteria. Putrescine is assumed to be the major biogenic amine associated with microbial food spoilage. Gram-negative bacteria can form putrescine by three metabolic pathways that can include eight different enzymes. The objective of this study was to design new sets of primers able to detect all important enzymes involved in the production of putrescine by Gram-negative bacteria. Seven new sets of consensual primers based on gene sequences of different bacteria were designed and used for detection of the speA, adiA, adi, speB, aguA, speC, and speF genes. A newly developed touchdown polymerase chain reaction (PCR) method using these primers was successfully applied on several putrescine-producers. Selected PCR products were sequenced and high similarity of their sequences (99-91%) with known sequences of the corresponding genes confirmed high specificity of the developed sets of primers. Furthermore, all the investigated bacteria produced both putrescine and agmatine, an intermediate of putrescine production, which was confirmed by chemical analysis. The developed new touchdown PCR method could easily be used to detect potential foodborne Gram-negative producers of putrescine. The newly developed sets of primers could also be useful in further research on putrescine metabolism in contaminating microbiota. © 2012 Elsevier Ltd. en
utb.faculty Faculty of Technology
dc.identifier.uri http://hdl.handle.net/10563/1003194
utb.identifier.obdid 43870139
utb.identifier.scopus 2-s2.0-84875664171
utb.identifier.wok 000317633600004
utb.identifier.coden FOMIE
utb.source j-scopus
dc.date.accessioned 2013-04-29T12:12:12Z
dc.date.available 2013-04-29T12:12:12Z
utb.contributor.internalauthor Wunderlichová, Leona
utb.contributor.internalauthor Buňková, Leona
utb.contributor.internalauthor Koutný, Marek
utb.contributor.internalauthor Valenta, Tomáš
utb.contributor.internalauthor Buňka, František
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