Risk-based Identification of Spoilage Bacteria for Quality Professionals
Empowering proactive decision-making for your brewery
Synergizing DNA Signature Capturing TechnologyTM and RT-PCR
Veripro features a multiplexed configuration that simultaneously detects multiple genetic targets, including an internal amplification control, in a single, pre-aliquoted tube. Results can be monitored in real-time to detect the presence and relative abundance of contaminants. A simplified data analysis approach enables clear interpretation of results, including spoilage potential, to empower rapid and effective quality decisions. Veripro maintains the same robustness, speed to results, and ease of use of the Veriflow platform, but leverages RT-PCR’s ability to multiplex and takes advantage of the platform’s capacity for high throughput analyses.
Delivering functional genetic analysis for the most critical beer spoilers
Our passion for innovation and the need for rapid, multiplexed, high-throughput assays for risk-based identification of beer spoilers while maintaining the ease-of-use and cost-effectiveness required by progressive breweries, led us to develop brewPRO for RT-PCR instruments.
The brewPRO Bacteria Screen assay provides risk-based detection and genetic characterization of Lactobacillus & Pediococcus species commonly found in beer. This information enables an assessment of spoilage risk based on the relative abundance of genes that confer resistance to inhibitory hops compounds. It also simultaneously detects the strictly anaerobic class of Megasphaera & Pectinatus species that can be near impossible to culture via traditional micro and severely affect products with lower levels of dissolved oxygen, ABV, and IBU.
The multiplexed assay simultaneously detects validated Veriflow gene targets in a single assay:
- Presence of Lactobacillus and Pediococcus species on the FAM channel
- Presence of hop resistance plasmids horA and horC that enable persistence and beer spoilage on the ROX channel
- Internal amplification control (IAC) to demonstrate successful amplification on the HEX channel
- Presence of Megasphaera and Pectinatus species on the Cy5 channel
Scientific approach provides unique information to assess spoilage risk
The discovery and characterization of hop resistance genes demonstrated that members of a single Lactobacillus or Pediococcus species vary widely in their ability to cause beer spoilage. Two hop resistant related genes known as horA and horC have been shown to enable lactic acid bacteria, such as Lactobacillus spp and Pediococcus spp, to grow in beer. The function of horA and horC is to encode transporters on the surface of cells that expel toxic hops compounds to enable survival in the presence of hops. These two genes were found to have nearly 100% correlation with species of beer spoiling lactic acid bacteria in several studies.1
However, mere detection of hop resistance genes is insufficient to fully understand the risk and potential for spoilage these organisms present. brewPRO reports BOTH the presence of horA and/or horC AND the relative abundance of these gene targets. This information serves as an indicator of bacterial persistence in beer and spoilage potential.
The graph below highlights the value of this information by comparing the relative abundance of several brewery isolates of Lactobacillus species. L. brevis is the predominant beer spoiler among Lactobacillus species, and its detection can strike immediate fear into brewers. However, not all L. brevis isolates are created equal. Several L. brevis isolates were tested with brewPRO and results demonstrate that each isolate poses a significantly different spoilage risk in beer. An isolate of L. plantarum, a subspecies not typically associated with beer spoilage, was also tested and results showed hop resistance capability and a relative abundance score that would make it a cause for concern in certain beer styles.
The higher the relative abundance score the higher the risk of spoilage.
Assessment of spoilage risk should also take into consideration beer style.
With this comprehensive information on spoilage potential, brewers can make more accurate decisions, and remediation activities can be tailored – improving overall beer quality and operational efficiencies. This unique information empowers quality teams to make real-time decisions about their processes by providing data on the specific nature of the organism in their facility from pitch through packaging.
Benefits of using brewPRO Bacteria Screen
- Superior, risk-based identification of the presence of Lactobacillus and Pediococcus in samples ranging from environmental to raw materials to in-process beer through finished products
- Proprietary analysis that quantifies the relative abundance of hop resistance genes
- Results in under 3 hours for wide variety of sample types
- Automated data collection and analysis
Unrivaled Ease of Use
- Simplified workflow with easy sample preparation - no DNA purification
- Multiplexed format
- High throughput with fast time to results
- Easy to train personnel
- Compatible with multiple instruments
- Ability to simultaneously analyze bacteria and wild yeast assays on a universal PCR amplification method
1 Suzuki et al, “A review of hop resistance in beer spoilage lactic acid bacteria”, Journal of the Institute of Brewing, 2006.
brewPRO Same Day Workflow
Adaptive workflow will accommodate colony pick, fermentation and yeast slurry as well as enrichment if desired
|brewPRO Bacteria Screen Performance Specifications|
|Sensitivity (LOD)||≤ 10 cells/ml or ≤ 1 cell per 100ml with enrichment|
|Time to Results||< 3 hours or 24-48 hours with enrichment|
|Matrix Compatibility||Beer, colony PCR, environmental, fermentation, yeast slurry & post-pasteurization (Viability application)|
|Assay Configuration||Multiplex, qualitative and risk assessment|
|Target Selection||Lactobacillus and Pediococcus-specific genomic target (FAM)|
Lactobacillus and Pediococcus-specific hop resistance genes horA and horC (ROX)
Megasphaera and Pectinatus-specific genomic target (Cy5)
Internal Amplification Control (HEX)
|Specificity||Lactobacillus species Including: L. brevis, L. buchneri, L. rhamnosus, L. jensenii, L. backii, L. paracollinoides, L. lindneri, L. delbrueckii, L. fructivorans, L. plantarum, L. pentosus, L. acetotolerans|
|Pediococcus species Including: P. damnosus, P. inopinatus, P. parvulus, P. pentosaceus, P. acidilactici, P. claussenii|
|Megasphaera species including: Megasphaera cerevisiae, Megasphaera paucivorans, Megasphaera sueciensis|
|Pectinatus species including: Pectinatus frisingensis, Pectinatus cerevisiiphilus, Pectinatus haikarae|