Your browser doesn't support javascript.
loading
Montrer: 20 | 50 | 100
Résultats 1 - 1 de 1
Filtre
Ajouter des filtres








Gamme d'année
1.
Braz. j. microbiol ; 48(4): 740-746, Oct.-Dec. 2017. tab, graf
Article Dans Anglais | LILACS | ID: biblio-889173

Résumé

ABSTRACT The spoilage of beer by bacteria is of great concern to the brewer as this can lead to turbidity and abnormal flavors. The polymerase chain reaction (PCR) method for detection of beer-spoilage bacteria is highly specific and provides results much faster than traditional microbiology techniques. However, one of the drawbacks is the inability to differentiate between live and dead cells. In this paper, the combination of propidium monoazide (PMA) pretreatment and conventional PCR had been described. The established PMA-PCR identified beer spoilage Lactobacillus brevis based not on their identity, but on the presence of horA gene which we show to be highly correlated with the ability of beer spoilage LAB to grow in beer. The results suggested that the use of 30 µg/mL or less of PMA did not inhibit the PCR amplification of DNA derived from viable L. brevis cells. The minimum amount of PMA to completely inhibit the PCR amplification of DNA derived from dead L. brevis cells was 2.0 µg/mL. The detection limit of PMA-PCR assay described here was found to be 10 colony forming units (CFU)/reaction for the horA gene. Moreover, the horA-specific PMA-PCR assays were subjected to 18 reference isolates, representing 100% specificity with no false positive amplification observed. Overall the use of horA-specific PMA-PCR allows for a substantial reduction in the time required for detection of potential beer spoilage L. brevis and efficiently differentiates between viable and nonviable cells.


Sujets)
Coloration et marquage/méthodes , Bière/microbiologie , Levilactobacillus brevis/isolement et purification , Levilactobacillus brevis/croissance et développement , Réaction de polymérisation en chaine en temps réel/méthodes , Propidium/analogues et dérivés , Propidium/composition chimique , Azotures/composition chimique , Levilactobacillus brevis/génétique , Levilactobacillus brevis/composition chimique , Réaction de polymérisation en chaine en temps réel/instrumentation , Microbiologie alimentaire
SÉLECTION CITATIONS
Détails de la recherche