Get results early. Rapidcult E. coli Enrichment Broth reduces your time to result for EHEC testing significantly.

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1 Get results early. Enrichment Broth reduces your time to result for EHEC testing significantly.

2 Enrichment Broth: Good things take time, very good things take less time. Time is crucial for sensitive food products. Quality is critical to protect your brand. But time and quality often conflict, especially when it comes to testing contamination in the production of meat and vegetables. It s time-consuming and slows your workflow, meaning the food is in quarantine and cannot be used. The Rapidcult TM E. coli Enrichment Broth is a medium for enriching pathogenic E. coli including E. coli O157 from food samples, especially from ground beef and beef trim. It reconciles scarce time with high reliability. This way, we help you speed up your workflow. s validation is consistent with USDA-FSIS when used in combination with our foodproof E. coli O157 real-time PCR kits or Singlepath E. coli O157 lateral flow tests. Questions about the use of the Merck Millipore test kit by USDA-FSIS-regulated facilities should be directed to AskFSIS ( Major advantage: Enrichment Broth offers detection after as little as 8-12 hours, but also allows you to incubate up to 22 hours to fit with your workflow. This saves you crucial time considering that classical enrichment media for E. coli can be read off after 24 hours at the earliest. Even with modern versions of E. coli media, time to result is 18 hours. This makes Enrichment Broth a faster and more flexible solution compared to other enrich ment media. In consequence, food products can be released much quicker and reach the customer fresher. So in the end not only saves you time, but also saves you money. E. coli O157 is not the only pathogenic E. coli. Other verotoxin-producing E. coli (EHEC) are dangerous as well. Initial results showed that with Rapidcult E. coli Enrichment Broth you are able to enrich other serotypes of pathogenic E. coli from different food sample materials, such as vegetables, apple juice or milk and diary products. Need quick information? Find out more about Enrichment Broth and how you can reduce your time to result for EHEC testing at Need extensive information? Contact your sales re presentative who will assist you with any questions. As you might have guessed: here, we re fast, too. 2

3 Find out how Enrichment Broth becomes a catalyst and not an obstacle to your workflow: 8-12 hours fit with your optimal workflow Shorter times to result lead to shorter times of food quarantine. Food reaches customer fresher Increased growth of E. coli due to special composition Comprehensive inclusivity study confirmed superior growth of all E.coli strains tested. Quality control process meets ISO guidelines. Granulated for maximum safety and convenience. Test for the best with at 3

4 Preparation protocol Enrichment Broth Protocol for 375 g sample Dissolve 43.8 g of dehydrated medium in 1.5 liters of pre-warmed (42 C), sterilized, deionized water Add 375 g of sample to pre-warmed (42 C) medium and homogenize Incubate 8 22 hours at 42 C or or Prepare immunoconcentrated solution of E. coli O157 by IMS Dispense 160 μl of the enriched sample into the sample port on Singlepath E. coli O157 Extract DNA from enriched sample using the foodproof StarPrep One Kit Streak onto CT-SMAC Read result after 20 minutes Prepare the reaction mix for the foodproof E. coli O157 Detection Kit Incubate 24 hours at C Confirm presumptive colonies on e.g. Fluorocult E. coli O157 4

5 Enrichment Broth Protocol for 25 g sample Dissolve 6.6 g of dehydrated medium in 225 ml of pre-warmed (42 C), sterilized, deionized water Add 25 g of sample to pre-warmed (42 C) medium and homogenize Incubate 8 22 hours at 42 C or or Prepare immunoconcentrated solution of E. coli O157 by IMS Dispense 160 μl of the enriched sample into the sample port on Singlepath E. coli O157 Extract DNA from enriched sample using the foodproof StarPrep One Kit Streak onto CT-SMAC Read result after 20 minutes Conduct RT-PCR analysis using the foodproof E. coli O157 Detection Kit Incubate 24 hours at C Confirm presumptive colonies on e.g. Fluorocult E. coli O157 5

6 Ordering information for testing of E. coli O157 After enrichment of the E. coli you can choose different microbiological methods for detection depending on the desired time to result. Merck Millipore s Enrichment Broth is perfectly optimized to the requirements of the following methods and reduces the risk of false negative results. Rapid detection with foodproof real-time PCR Enrichment of E. coli Sample preparation StarPrep One 96 tests ShortPrep II Kit 96 tests PCR foodproof E. coli O157 5 Nuclease 96 tests foodproof E. coli O157 Hybridization probes (LC1.x, 2.0) 96 tests Rapid screening with Singlepath E. coli O157 Enrichment Singlepath E. coli O tests Classical culture method Enrichment of E. coli Detection of E. coli O157 Sorbitol-MacConkey Agar (SMAC Agar) 500 g CT-Supplement 10 vials Confirmation Fluorocult E. coli O157: H7 Agar 500 g piece Singlepath E. coli O tests

7 Ordering information for testing of EHEC In addition to enrichment of E.coli O157, Rapidcult can be used for testing for the presence of verotoxinproducing E. coli such as E. coli O104 and other EHEC in various sample types (meat, milk and dairy products, apple juice, fruit and vegetable products etc.). Rapid method for screening with Duopath Verotoxins Enrichment of E. coli Post-enrichment CAYE Broth 100 g CAYE Broth Supplement 10 vials Polymyxin B sulfate Bacillus cereus selective supplement Screening 10 vials tests Classical microbiological method Enrichment Detection Chromocult Coliform Agar 500 g Chromocult TBX Agar 500 g Confirmation Post-enrichment CAYE Broth 100 g CAYE Broth Supplement 10 vials Polymyxcin B sulfate Bacillus cereus selective supplement 10 vials tests

8 For further information on Merck Millipore and our products contact: Merck KGaA Merck Millipore Darmstadt, Germany foodproof E.coli 0157 Detection Kit Hybridization Probes (LC1.x, 2.0) License Notice: The purchase of this product includes a limited, non-transferable license under US Patents Nos. 6,245,514 and 6,174,670, and corresponding patents and patent applications outside the United States, owned by the University of Utah Research Foundation and licensed by Idaho Technology, Inc. and Roche Diagnostics GmbH, to use only this amount of product for FRET assays solely for food analysis or GMO analysis. No right is conveyed, expressly, by implication or estoppel, for any other patent, such as under any patent for an apparatus or system, or to use this product for any other purpose. Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 6,127,155, 5,677,152 (claims 1 to 23 only), and 5,773,258 (claims 1 and 6 only), and claims outside the US corresponding to expired US Patent No. 5,079,352. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product solely in Food Testing Applications and Genetically Modified Organism (GMO) Testing Applications, including reporting results of purchaser s activities for a fee or other commercial consideration, and also for the purchaser s own internal research. No right under any other patent claim is conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. The purchase price of this product includes limited, nontransferable rights under US Patent No. 7,687,247 owned by Life Technologies Corporation to use only this amount of the product to practice the claims in said patent solely for activities of the purchaser for bioburden testing, environmental testing, food testing, or testing for genetically modified organisms (GMO) in accordance with the instructions for use accompanying this product. No other rights are conveyed, including no right to use this product for in vitro diagnostic, therapeutic, or prophylactic purposes. Further information on purchasing licenses under the above patent may be obtained by contacting the Licensing Department, Life Technologies Corporation, foodproof E. coli 0157 Detection Kit, 5 Nuclease License Notice: Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,804,375, 5,538,848, 5,723,591, 5,876,930, 6,030,787, 6,258,569. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product solely in Food Testing Applications and Genetically Modified Organism (GMO) Testing Applications, including reporting results of purchaser s activities for a fee or other commercial consideration, and also for the purchaser s own internal research. No right under any other patent claim is conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. The purchase price of this product includes limited, non-transferable rights under US Patent No. 7,687,247 owned by Life Technologies Corporation to use only this amount of the product to practice the claims in said patent solely for activities of the purchaser for bioburden testing, environmental testing, food testing, or testing for genetically modified organisms (GMO) in accordance with the instructions for use accompanying this product. No other rights are conveyed, including no right to use this product for in vitro diagnostic, therapeutic, or prophylactic purposes. Further information on purchasing outlicensing@lifetech.com or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking the suitability of our products for the envisaged purpose. The M mark, Chromocult, Duopath, Fluorocult and Singlepath are registered trademarks of Merck KGaA, Darmstadt, Germany. foodproof is a trademark of Biotecon Diagnostics GmbH.