This product is developed for the identification and quantification of yeasts belonging to the Brettanomyces/Dekkera genus from filterable and non-filterable beverage products (juices, teas, soft drinks, energy drinks, concentrates and wine) The name Brettanomyces is used for the non-spore forming form of the yeast while Dekkera refers to the telemorph form. The genus is in the family of Saccharomycetaceae and includes five known species: B/D. bruxellensis, B/D. anomalus/anomala, B/D. custersianus/custersiana, B/D. nanus/nana, B/D. naardenensis. The yeast is acid-producing, when grown on glucose rich media under aerobic conditions, produces large amounts of acetic acid. Therefore, it is currently considered as a spoilage organism responsible for off-flavor production in different products such as wine, cider, beverages or dairy products. In nature, Brettanomyces can be found on the skins of fruit. It is tolerant to low pH, growth conditions show that it prefers acidic environments and room temperature but thermal inactivation is usually observed only at temperatures over 50°C. The system is based on the amplification of DNA region selectively from Brettanomyces/Dekkera and signal detection of probes constructed as hybridisation probes for the amplified product. The DEK LC RT-PCR Workflow System has been designed based on NCBI (National Center for Biotechnology Information) nucleotide database. It was blasted and tested against bacteria, moulds and yeasts other than Brettanomyces/Dekkera. In these cases no signal was detected. The DEK LC RT-PCR Workflow System Kit includes:
- DEK Pre-cultivation Kit
- DEK Isolation Kit
- Brettanomyces/Dekkera detection and identification Real-Time PCR Kit
DEK LC RT-PCR Workflow System uses automated nucleic acid purification, automated PCR set up and automated detection in results reporting. The groups bellow can be identified by the DEK LC RT-PCR Workflow System based on the melting temperature of the probes:
|Name of microorganisms||Melting temperature data, C°||Number of type strains|
|B/D.bruxellensis or B/D.anomalus/anomala||52,8||DSM70001 DSM70727|
|B/D. nanus/ nana||44,4||ATCC48014|
Fig 1: Identification of Brettanomyces/Dekkera yeasts by DEK LC RT-PCR Workflow
- System The DEK LC RT-PCR Workflow System advantages:Selective detection and identification of viable Brettanomyces/Dekkera on species level. The detection limit is 1 cfu/200 ml product.
- Quantification of Brettanomyces/Dekkera yeasts is solved.
- System gives information on the status of cell culturability.
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