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Statements

Subject Item
n2:RIV%2F70883521%3A28110%2F13%3A43869930%21RIV14-GA0-28110___
rdf:type
skos:Concept n19:Vysledek
rdfs:seeAlso
http://ac.els-cdn.com/S0740002012002341/1-s2.0-S0740002012002341-main.pdf?_tid=906ed560-acad-11e2-b4d2-00000aacb35e&acdnat=1366787403_27598ec11c61607000a2e1f0584334af
dcterms:description
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. 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.
dcterms:title
Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products
skos:prefLabel
Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products Novel touchdown-PCR method for the detection of putrescine producing Gram-negative bacteria in food products
skos:notation
RIV/70883521:28110/13:43869930!RIV14-GA0-28110___
n19:predkladatel
n20:orjk%3A28110
n4:aktivita
n21:S n21:P
n4:aktivity
P(GAP108/10/0200), P(GAP503/11/1417), S
n4:cisloPeriodika
2
n4:dodaniDat
n18:2014
n4:domaciTvurceVysledku
n9:3269531 n9:6731007 n9:8264945 n9:2594862
n4:druhVysledku
n6:J
n4:duvernostUdaju
n16:S
n4:entitaPredkladatele
n10:predkladatel
n4:idSjednocenehoVysledku
92385
n4:idVysledku
RIV/70883521:28110/13:43869930
n4:jazykVysledku
n8:eng
n4:klicovaSlova
Agmatine, Gram-negative bacteria, Multiplex PCR, PCR, Primers, Putrescine
n4:klicoveSlovo
n7:Multiplex%20PCR n7:Gram-negative%20bacteria n7:Agmatine n7:Primers n7:Putrescine n7:PCR
n4:kodStatuVydavatele
GB - Spojené království Velké Británie a Severního Irska
n4:kontrolniKodProRIV
[0C8383800542]
n4:nazevZdroje
Food Microbiology
n4:obor
n5:GM
n4:pocetDomacichTvurcuVysledku
4
n4:pocetTvurcuVysledku
5
n4:projekt
n11:GAP108%2F10%2F0200 n11:GAP503%2F11%2F1417
n4:rokUplatneniVysledku
n18:2013
n4:svazekPeriodika
34
n4:tvurceVysledku
Koutný, Marek Wunderlichová, Leona Valenta, Tomáš Buňka, František Buňková, Leona
n4:wos
000317633600004
s:issn
0740-0020
s:numberOfPages
9
n17:doi
10.1016/j.fm.2012.09.024
n12:organizacniJednotka
28110