描述

The PIKA Weihenstephan™ SO Detection Kit B LP Identification enables rapid identification of 10 Lactobacillus species and Pediococcus. Typically used after obtaining a positive result using the PIKA Weihenstephan™ SO Detection Kit B LP Screening, the PIKA Weihenstephan™ SO Detection Kit B LP Identification identifies the individual presence or absence of all the main lactic acid beer spoiling bacteria including Lactobacillus backii, L. brevis, L. casei, L. collinoides, L. coryniformis, L. parabuchneri (“frigidus"), L. lindneri, L. perolens, L. plantarum, and L. rossiae by real-time PCR. It also identifies Pediococcus damnosus/inopinatus. These twelve organisms represent the most common spoilage bacteria in beer and breweries. The kit includes reagents for both DNA sample preparation and real-time PCR-based detection and yields reliable results from enriched beer and brewery samples usually in ~2 hours. Because analysis is performed directly on enriched samples, there is no need to isolate bacteria to single colonies, saving time and yielding results days faster than culture-based methods.

• Fast—from enriched beer samples to results usually in ~2 hours
• Sensitive—detects as few as 1000 cells per sample, enabling detection long before visible growth
• Simple—pre-aliquotted, ready-to-use format minimizes errors and pipetting steps. Sample prep reagents included.
• Stable—shelf-stable reagents can be stored at room temperature

How the Kit Works
After enrichment of beer or brewery samples in an appropriate media such as PIKA Weihenstephan™ FastOrange™ B Broth* (supplied separately) and rapid DNA sample preparation using the reagents supplied in the kit, detection is accomplished by real-time PCR using the included TaqMan® assay reagents. Each sample is tested in 11 separate reactions for Lactobacillus backii, L. brevis, L. casei, L. collinoides, L. coryniformis, L. parabuchneri (“frigidus"), L. lindneri, L. perolens, L. plantarum, L. rossiae, and Pediococcus damnosus/inopinatus.

Because of its exquisite sensitivity and specificity, real-time PCR is an ideal method for detecting beer spoilage bacteria within mixed cultures, even among high concentrations of brewer’s yeast. The spoilage organisms can often be detected from clear samples before the culture is visibly turbid, so results can be obtained days earlier than with traditional culture methods. Plating to individual colonies is unnecessary, and there is no guesswork regarding the presence of any of the target species of beer spoilage organisms.

The TaqMan® primer/probe sets, which are included in individual ready-to-use reaction tubes, target DNA sequences of the most common beer spoilage bacteria, including Lactobacillus backii, L. brevis, L. casei, L. collinoides, L. coryniformis, L. frigidus, L. lindneri, L. perolens, L. plantarum, L. rossiae, Pediococcus damnosus, and P. iopinatus. Upon amplification, the target sequences are detected in the FAM™ fluorescence channel of the real-time PCR instrument. An internal positive control (IPC) included in the Pediococcus assay is detected in the VIC® channel. The IPC is included to verify the absence of PCR inhibitors and that amplification proceeds normally in the reaction.

*We strongly recommend the use of PIKA Weihenstephan™ FastOrange™ B Broth to enrich beer or brewery samples prior to analysis with the PIKA Weihenstephan™ SO Detection Kit B LP Identification. FastOrange™ B Broth is compatible with all types of beer samples, as well as other brewery samples such as water and pitching yeast. It is optimized for the rapid and selective growth of beer spoilage bacteria and has been qualified for use with downstream PCR applications.

Workflow
Beer or brewery samples—even those with high yeast concentrations—are typically enriched in an equal volume of ready-to-use PIKA Weihenstephan™ FastOrange™ B Broth for 2–3 days at 25°C, then sample preparation is accomplished using a simple heat lysis method. Briefly, 1 mL of the enriched sample is pelleted by centrifugation, optionally washed, and then subjected to a 10-minute high temperature lysis using the included wash and lysis reagents. After a brief centrifugation step, the lysate is ready for analysis by PCR.

PCR set up is made easy with the pre-aliquotted primer-probe mix. Simply add the included Rehydration Buffer, TaqMan® Universal Master Mix II-No UNG (available separately, Cat. No. 4440043), and prepared DNA sample to the supplied individual reaction tubes of dried primer-probe mix. Real-time PCR is then performed for 40 cycles. The entire process from DNA sample preparation to results takes only about two hours, with very little hands-on time.

The kit is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, and 7500 Fast real-time PCR systems.

Backed by Experts
The PIKA Weihenstephan™ SO Detection Kit B LP Identification was developed by and is manufactured for Life Technologies by PIKA Weihenstephan GmbH. Since 2000, PIKA Weihenstephan has been a consultant and leading provider of testing products and services for the brewing and beverage industry.

规格

Target Organism:	L. lindneri, L. plantarum, L. collinoides, L. backii, P. inopinatus., L. parabuchneri, L. perolens, L. casei, L. rossiae, L. coryniformis, L. brevis, P. damnosus
Validated Application:	Real-Time PCR
Detection Method:	Primer-Probe Detection
Platform:	7500 Fast System, StepOnePlus™ System, StepOne™ System
Sample Type (Specific):	Enriched Beer Samples, Brewery Samples
Product Size:	8 x 11 reactions

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