Microbiological testing is critical at every stage of beverage production, such as manufacturing, winemaking or brewing, to maintain product quality. Rapid microbiological analysis techniques such as qPCR can improve the reliability of spoilage detection in beverages while reducing labour costs and product delays. However, these methods are of limited use because of small analytical volumes. To address this problem, overnight pre-enrichment is often used, but this negates the time-saving benefit of rapid analysis.
InnovaPrep has developed a rapid "concentration" method to eliminate the need for culture-based enrichment steps. This technique uses a single-use high-flow membrane filter and an innovative elution process.
The InnovaPrep concentration pipette system rapidly concentrates microorganisms, up to a concentration of 10,000 times, from liquid samples. The system filters the sample and then uses a wet foam elution method to rapidly recover microorganisms in a much smaller volume. The system's ease of use and ability to provide exceptionally high concentration factors and simultaneous clean buffer exchange make this system an ideal solution for trace analysis of spoilage organisms.
The concentrate pipette system is suitable for bacteria, fungi, spores and viruses.
Required materials
• Instrument: CP Select and CP Select User Guide
• InnovaPrep Be Flat™ Degassing Jar Item #HC07674
• Consumables:
- Elution Buffer:
- Concentrating Pipette Tips (CPTs):
- 0.45 µm CPTs
Procedure
STEP 1 - Choose elution buffer.
Choose from the following elution buffer formulas depending on your method of analysis:
- Classic culture- HC0800- 0.075% Tween 20 PBS
- Rapid methods- HC08001- 0.075% Tween 20 Tris
STEP 2 - Sample decontamination
It is necessary to decarbonate beverages before processing on the Concentration Pipette because CO₂ in the liquid interferes with the flow sensor and tends to blind the membrane. InnovaPrep has developed the Be Flat™ Degassing Pot, which has a unique, patented design to quickly degass carbonated beverages.
To degas, you quickly pour the beverage at room temperature into the Be Flat™ Degassing Pot from a few inches above the membrane. This will release as much CO₂ as possible. Place the jar in the refrigerator for 10 minutes.
Note: It is important to evaluate the optimal time for degassing as each beverage is different. Some beverages do not require a cooling step, while others may take up to 20 minutes to degas.
STEP 3 - Concentration
- Set up the CP Select according to the instructions in Chapter 4 of the User's Manual.
- Insert a CPT and select the HOLLOW protocol from the menu.
- Lower the probe into the sample.
- Press "Start Run" on the user screen. When the entire sample has been processed, the CP stops.
- Place a clean final sample container under the CPT. The menu screen prompts you to press "Elute".
- Press "Elute." The sample is dispensed from the pipette tip into the sample container. The sample is ready for next steps for sample preparation and analysis.
Note: Process volumes vary depending on beverage type, temperature and the amount of residual carbon dioxide in the sample.
Method of analysis
- Samples are suitable for a variety of culture and rapid methods, including digital PCR/qPCR, digital RT-PCR/RTqPCR, NGS, Lateral Flow Assays, etc. The samples concentrated by the CPT are now ready for analysis and any further downstream processing steps.
- Store pre-concentrated or post-concentrated liquid samples at 4 °C.
