Food Safety Testing Solutions by QIAGEN Brochure

Food Safety Testing Solutions by QIAGENSample & Assay TechnologiesCatch it real-timeA fully integrated system for comprehensive food safety testing2 www.qiagen.com Food Safety Testing 10/2010In a globalized food market with increasing demand for food research and monitoring, there is a need for streamlined testing solutions that are sensitive, accurate, and easy to use with a variety of starting materials. Real-time polymerase chain reaction (PCR) is gaining importance as a complementary tool to established microbial culture-based methods and ELISA-based assays prevalent in the food testing sector. Combined with dedicated sample preparation kits, real-time PCR assays deliver rapid results from a broad spectrum of sample materials and simplify the testing process.Meeting the challenges of global food distribution¦n Real-time PCR delivers results in approximately 3 hours¦n Real-time PCR accurately detects challenging pathogen strains¦n Protocols are simple and can be automated for high sample numbers¦n Real-time PCR is highly specific and unaffected by food processingTop performance Assays must be sensitive, specific for the tested target, and resistant to inhibition of detection. This performance is dependent on the quality of input material and thus, sample preparation methods must be effective.Ease of use Both sample preparation methods and detection assays must be based on streamlined, robust, and user-friendly protocols that are easily adapted to varied applications.Speed Sample preparation must involve minimal effort and, if possible, should be automatable to process high sample numbers. The detection assays must deliver rapid results and accommodate high throughput.Coverage Sample preparation methods must accommodate a broad range of sample types while delivering efficient extraction with a streamlined protocol. In addition, the detection assays must cover a wide range of known food contaminants or pathogens.StandardizationBoth sample preparation and detection assays must integrate into a consistent workflow, allowing standardization of sample processing.A modern method for food testingClear advantages account for the popularity of molecular-based testing:IntroductionToday’s high-quality food safety testing program places demands on testing assays, as well as sample preparation methods:An integrated testing system that meets your demandsUniversal sample preparationFood Safety Testing 10/2010 Sample & Assay Technologies 3The mericon™ food testing portfolio is a complete system of assay kits that meet the demands listed above. Based on detection by real-time PCR, mericon PCR Assays and sample preparation kits enable fast and reliable detection of a broad range of pathogens, genetically modified organisms, allergens, and plant and animal matter in food, animal feed, or pharmaceutical products. A major advantage of the QIAGEN setup is harmonized detection protocols across the portfolio, resulting in workflows that differ only in sample preparation method (Figure 1). Complementing the aligned PCR-based detection assays, QIAGEN offers new sample preparation solutions specifically designed to process a variety of food materials, providing an optimal overall workflow performance. These sample preparation kits efficiently extract DNA from microbial enrichment cultures as well as directly from various food matrices, while minimizing the carryover of PCR inhibitors inherent to complex food samples. Sample preparation that is universal, simplified, and rapidOur sample preparation kits are designed to enable DNA extraction from diverse sample types, starting either from a microbial enrichment culture or from raw or processed food material. Particularly noteworthy is the improved CTAB-based extraction method of the DNeasy® mericon Food Kit. The nonionic detergent CTAB is widely used for efficient extraction of total cellular nucleic acids from a wide range of tissue types. CTAB can be adjusted to either efficiently liberate and complex with DNA or to remove cellular inhibitors. Compared to conventional, labor-intensive CTAB extraction protocols, the protocol of the DNeasy mericon Food Kit is significantly faster, accomplishing up to 30 extractions in 2.5 hours with a radically reduced number of workflow steps (Figure 2). The extracted DNA is of equal quality and purity as that extracted with conventional CTAB protocols.Figure 1. A two-armed workflow for food safety testing. Whether testing for pathogens, genetically modified organisms, allergens, or animal and plant matter, the mericon food testing portfolio delivers solutions for a streamlined sample preparation workflow and a fully standardized detection protocol.Suspension in CTAB extraction solutionSuspension in CTAB lysis buffer60 min incubationat 65°C30 min incubation with Proteinase K Solution at 65°CFirst chloroform extraction Chloroform extractionSecond chloroform extraction (aqueous phase)Addition of binding buffer to aqueous phaseDNA precipitation by incubation at 65°CCentrifugation and resuspension of DNA pelletDNA precipitation with isopropanolAddition of CTAB precipitation solutionPurification of DNA with spin-column technology30 samples processed in 1 workday30 samples processed in 2.5 hConventional CTAB food lysisQIAGEN CTAB food lysismericon DNA Bacteria Plus KitDNA extraction from Gram-positive bacteriamericon DNA Bacteria KitDNA extraction from Gram-negative bacteria DNeasy mericon Food KitDNA extraction from all food typesMicrobial enrichment culturemericonPCR Assays Detection by real-time PCRTemplate DNAFood SampleFigure 2. Efficient and rapid DNA extraction. The DNeasy mericon Food Kit enables DNA extraction from all food samples in radically shorter time compared to conventional CTAB protocols and with high recovery of short DNA fragments. Times given include handling and instrument time.Key features that simplify food safety testing¦Fast and sensitive detection4 www.qiagen.com Food Safety Testing 10/2010During production, food is subjected to heat, irradiation, high pressure, and changes in pH which lead to DNA degradation and fragmentation. The optimized chemistry of the DNeasy mericon Food Kit was developed to recover short DNA fragments (down to 100 bp) ensuring that even highly fragmented DNA is efficiently isolated and subsequently amplified in PCR. With these features, extraction with the DNeasy mericon Food Kit is a universally applicable extraction method that generates optimal and reliable results even when using strongly inhibitory, highly processed, fatty, acidic, high, or low DNA content foods (Figures 3 and 4). In addition, establishing multiple lysis procedures for different food matrices is no longer necessary. Short time to result and adaptable throughputTesting of a food sample, from sample preparation and assay setup to detection by real-time PCR, takes approximately 3 hours. Furthermore, the simplicity of the protocols makes the workflow adaptable to application- or laboratory-specific throughput requirements, making even a high-throughput food safety testing program that covers multiple targets and sample types, feasible and efficient.Highly sensitive and specific detection of targetsReal-time PCR is a very sensitive and highly specific detection method. To further increase the sensitivity and specificity of mericon PCR Assays, the kits include Multiplex PCR Master Mix, which features highly efficient HotStarTaq Plus Polymerase, patented multiplex PCR technology such as Factor MP, and fast cycling technology including Q-bond®. These features create a reaction environment that is sensitive, specific, efficient, and highly resistant to PCR inhibitors found in food sample matrices (Figure 4). Generally, the sensitivity of each mericon PCR Assay is as little as 10target copies in a reaction. The specificity of each kit has been tested against large sets of non-target DNA from related species.Figure 3. High yields of DNA and minimal inhibitor carryover. Peanut butter and chocolate (difficult and highly inhibitory food matrices) were prepared using the standard protocol of the DNeasy mericon Food Kit and kits from 3 other suppliers to amplify a segment of the trnL chloroplast tRNA gene. nA Threshold cycle values (CT) are lower for samples prepared with the DNeasy mericon Food Kit compared to the other kits. nB Performed with pure DNA,this assay gives a positive CT shift of 2.9 – 3.5 when a sample is diluted 1:10. Lower shifts indicate the presence of PCR inhibitors. Samples were diluted 1:10 and the same DNA segment was amplified. Eluates prepared with the DNeasy mericon Food Kit were free of inhibitors, as evidenced by CT shifts of approximately 3. Eluates prepared with the other kits showed strong inhibitor contamination. All data were normalized to a fixed sample input to account for differences in kit input amount, dilution steps, and elution volume.* Only one replicate resulted in DNA amplification. 25.024.524.023.523.0 22.522.021.521.020.520.0Threshold cycle value (CT)APeanut butterDNeasy mericonFood KitSupplier A Supplier B Supplier C4543403835 333028252320Threshold cycle value (CT)DNeasy mericonFood KitSupplier A Supplier B Supplier CChocolateDNeasy mericonFood KitSupplier A Supplier B Supplier C 7531-- 1 -- 3-- 5-- 7-- 9--11Change in threshold cycle value (?C T)BPeanut butter Chocolate¦Standardized detection procedureFood Safety Testing 10/2010 Sample & Assay Technologies 5Figure 4. Heightened resistance to inhibitors. A plasmid carrying an artificial DNA sequence (DNA model TaqMan® System; 108 copies) was amplified in reactions spiked with inhibitorsand containing the Multiplex PCR Master Mix supplied with mericon PCR Assays or the mastermix provided by Supplier AB. Inhibitors and concentrations were chosen to simulate potential contaminations carried over from conventional sample preparation methods. Threshold cycle value of each reaction was compared to that of a negative control where water was added to the PCR. Reported is the change in threshold cycle value, which is higher for reactions performed with the master mix from Supplier AB, demonstrating the protective capacity of the Multiplex PCR Master Mix.Validated workflow solutions The entire workflow for food safety testing using mericon samplepreparation kits and mericon PCR Assays has been validated. mericon PCR Assays perform optimally on the Rotor-Gene® Q real-time PCR instrument, but have been validated for block thermal cyclers as well. The described analytical PCR procedure allows the customer to perform analysis according to local official requirements. In addition, mericon PCR Assays are fully licensed for use of PCR, eliminating any additional costs.Uniform and all-inclusive kit formatAll mericon real-time PCR kits contain the same components: target-specific primers and probes, an internal control to indicate possible PCR inhibition, a target-specific positive control, and all reagents necessary to perform the PCR assay. The real-time PCR protocol has been standardized across the portfolio so that the same cycling conditions are used for every target. Thus, a standardized detection procedure can be used for all targets within all testing segments and a variety of targets can be analyzed within a single thermal cycler run.Change in threshold cycle value (?C T)CTAB (0.04 mM) NaCI (50 mM) iPrOH (0.7 mM) EtOH (0.8 mM)14121086420Multiplex PCR Master MixSupplier ABPathogensSensitive detection of microbial contaminants in foodFigure 5. Comparison of time to result of different methods for pathogen detection. Delivering sensitive and highly specific results, real-time PCR analysis is also faster than conventional pathogen detection methods. Besides taking longer, immunoassays and traditional culture methods are less sensitive and more labor intensive. Figure 6. Highly sensitive pathogen detection, even in difficult food matrices such as peanut butter. Peanut butter was homogenized in buffered peptone water, spiked with < 5 cfu of Salmonella, and enriched for 20 h at 37°C. DNA was extracted from serially diluted samples of the enrichment culture using the mericon DNA Bacteria Kit and then tested with the mericon Salmonella Kit. Although the original inoculation was small, Salmonella was still reliably detected at a dilution factor of 1:100,000.6 www.qiagen.com Food Safety Testing 10/2010To date, the predominant method to test food for the presence of pathogens has been via culture preparation and/or ELISA-based assays. Molecular-based methods, specifically real-time PCR, offer an informative complement to these well-established methods. In addition, real-time PCR detection delivers results in less time (Figure 5), affords greater sensitivity and specificity, and has the robustness to cope with difficult food matrices (Figure 6). Finally, real-time PCR is a safe method for detecting challenging pathogen strains. Primary enrichmentSecondary enrichmentDetection by immunoassayTraditional culture: primary enrichmentSelectiveisolationConfi rmation of pathogenTraditional culture: secondary enrichmentPrimary enrichmentDNA extractionDetection by real-time PCR24 h 18 – 27 h 1.5 – 2 days 3 – 7 daysSensitive, specific, and reliable detection of pathogenic contaminantsDecreased specificityHighly labor intensiveNormalized fluorescence451:1000 dilution1:100 dilution1:10,000 dilution1:100,000 dilutionCyclePathogensReal-time PCR: a sophisticated tool to maximize specificityAs pathogenicity may differ among species and even strains of a particular microbial contaminant, at times it is necessary to reliably detect a single pathogen species or strain. Because real-time PCR detection is based on the amplification of very specific DNA sequences, mericon PCR Assays have the level of specificity needed to accurately distinguish, for example, the highly pathogenic Listeria monocytogenes from among other common Listeria species, or specifically detect shigatoxin-secreting Escherichia coli. Easy and reliable sample preparation To provide a streamlined and straightforward method for microbial sample preparation from pathogen enrichment cultures, QIAGEN has designed protocols based on mechanical lysis (using beads) and thermal lysis (by boiling) that deliver optimal disruption force for different bacteria types. Thermal lysis is optimized for the majority of easily lysed Gram-negative bacteria (e.g., Salmonella spp., Campylobacter spp., and Cronobacter spp.), whereas the advanced features of mechanical sample preparation create the lysis conditions needed for difficult Gram-positive bac-teria, such as Listeria spp. (Figure 7). Developed to be combined with mericon PCR Assays, both sample preparation systems deliver a DNA suspension that is optimal for real-time PCR analysis. Carefully formulated chemistry included in the mericon DNA Bacteria Kit and the mericonDNA Bacteria Plus Kit maximizes lysis efficiency, stabilizes the extracted DNA, and removes inhibitors that may interfere with downstream applications.With the methods of these dedicated sample preparation kits, DNA extracted from even low-count samples is still suitable for sensitive real-time PCR. The flexibility of the method, using either mechanical or thermal lysis, accommodates a range of microbial species. Figure 7. Efficient DNA extraction from Gram-negative or Gram-positive bacteria. nA Samples of an enrichment culture of Listeria monocytogenes were diluted and subjected to the mechanical (bead) lysis protocol of the mericon DNA Bacteria Plus Kit and the thermal (boiling) lysis protocol of the mericon DNA Bacteria Kit to evaluate efficiency of lysis. For this Gram-positive bacterium, mechanical lysis delivered a more effective extraction, most evident at very low bacterial concentrations (highest dilution) where the average threshold cycle value was 4.9 cycles lower with mechanical lysis. nB In a similar experiment, diluted and undiluted samples of an enrichment culture of Salmonella enterica in milk and hazelnut spread were subjected to the mechanical and thermal lysis protocols. Overall, the efficiency of DNA extraction from Gram-negative bacteria is similar with both preparation protocols.Food Safety Testing 10/2010 Sample & Assay Technologies 701020304050Threshold cycle value (CT)Mechanical lysisThermal lysis0510152025303535Threshold cycle value (CT)Undiluted1:101:1001:10001:10,0001:100,0001:1,000 000 Milk MilkHazelnut spreadHazelnut spreadUndiluted enrichment culture 1:10,000 diluted enrichment cultureMechanical lysisThermal lysisBAGenetically modified organismsDetermining the presence of genetically modified organisms (GMO) in food entails detection of genetic constructs that are used to generate transgenic and cisgenic organisms. Real-time PCR is the method of choice to test food for the presence of matter originating from GMOs, since this technology specifically detects the presence of target DNA sequences. mericon PCR Assays feature primers and probes that target frequently used genetic constructs, such as the nos terminator and the 35S promoter. As a result, these tests deliver a decisive detection signal in the presence of widely used GMOs (Table 1). For the identification of a particular GMO, the mericon food testing portfolio includes a suite of tests that target DNA sequences unique to each of a wide range of GMOs, such as Roundup Ready soy, Bt11 corn, or MON810 corn. Detection with these kits is highly sensitive and specific. Table 1. Results from cross-reactivity experiments for the mericon Screen Nos Kit and mericon Screen 35S Kit Speciesmericon Screen Nos Kitmericon Screen 35S KitBt11 corn + +Bt176 corn – +GA21 corn + –CaMV – +Barley – –Wheat – –Pig – –Horse – –CBH351 corn + +LL T25 corn – +Roundup Ready soy + +LL canola – +Potato – –Rye – –Sheep – –NK603 corn + +MON810 corn – +Roundup Ready canola – –Lambda DNA – –Rice – –Cattle – –Goat – –8 www.qiagen.com Food Safety Testing 10/2010Reliable identification of genetically modified organisms + : positive detection.– : no detection signal.Genetically modified organismsOne efficient sample preparation solution for all starting materialsThe samples used to test for genetically modified organisms are often raw plant matter, seeds, and processed foods, which are challenging starting materials consisting of difficult food matrices. The extraction of sufficient and high-quality DNA is essential for reliable detection of GMOs and key to the sensitivity of any testing system. The DNeasy mericon Food Kit addresses this need by enabling efficient extraction of DNA from a broad range of heterogeneous starting materials using a streamlined, universal CTAB-based DNA preparation protocol (page 3). With fewer workflow steps than conventional CTAB protocols, up to 30 samples can be processed in 2.5 hours. DNA recovery with these kits is demonstrably high and efficient, even with DNA fragments down to 100 bp (Figure 8). The DNeasy mericon Food Kit complements mericon PCR Assays for detecting GMOs creating a streamlined workflow for efficient sample processing and reliable results. Figure 8. Enhanced DNA fragment recovery with optimized protocols. Samples of cornflakes and ketchup, strongly processed foods which consequently contain highly fragmented DNA, and cocoa, an inhibitor-rich food matrix, were processed using the standard protocol and the small fragment protocol of the DNeasy mericon Food Kit. To test the recovery capacity of these protocols, the samples were spiked with human DNA fragments 100 bp long prior to purification on spin columns. Human DNA fragments in the eluate were amplified using a probe-based PCR system specific for the added fragment. Binding conditions of the small fragment protocol significantly improved recovery in all three samples, resulting in threshold cycle values that were lowerby up to 3.1 cycles (red, green, and pink curves) compared to the standard protocol (blue curves). This equates to approximately a 10-fold improvement in recovery of small fragments. Food Safety Testing 10/2010 Sample & Assay Technologies 90.10.20.30.40.55 10 15 20CycleCornflakes ? CT = 3.125 30 35 40Normalized fluorescence5 10 15 20CycleKetchup ? CT = 1.225 30 35 40Normalized fluorescence0.050.100.150.200.250.300.350.400.450.505 10 15 20CycleCocoa ? CT = 1.425 30 35 40Normalized fluorescence0.050.100.150.200.250.300.350.400.450.50 ELISA-based methods prevail in allergenic testing despite well-known conditions that lead to cross-reactivity (e.g., allergic cross-reactivity between almond and peanut). PCR-based methods are taking a stronghold in this area due to the superior specificity afforded by direct detection of allergen-specific DNA sequences. This heightened specificity practically eliminates concerns regarding testing cross-reactivity (Table 2). In addition, the sensitivity and specificity of allergen DNA detection by real-time PCR are independent of the ratio of allergen DNA to food DNA. This ratio often poses a problem for ELISA because allergens are typically present in trace amounts and thus, the immunological signal of a given allergen can be overwhelmed by cross-reactive molecules in the food matter. mericon PCR Assays detect allergen DNA when present in amounts as low as 10 copies per reaction. As a consequence, real-time PCR delivers highly specific detection of allergy-causing food fractions.Evidence-supported validity of allergen DNA detectionUltimately, the agent causing an allergic reaction is a protein. This raises the question whether basing allergen testing on molecular methods that screen for DNA is appropriate, especially when allergen quantification is a goal. There is ample evidence for a correlation between allergen quantification based on the direct detection of allergen protein versus the detection of allergen DNA (1).Allergens10 www.qiagen.com Food Safety Testing 10/2010Eliminating cross-reactivity from allergen testingTable 2. Assay specificity for walnut DNASpecies ResultWalnut +Peanut –Hazelnut –Almond –Pistachio –Pine nut –Pecan –Macademia –Brazil nut –Pumpkin seed –Sunflower seed –Sesame seed –Plum –Apricot –Cherry –Canola –Corn –Lentil –Lupine –Soy –Rye –Sorghum –Pea –Buckwheat –Celery – + : positive detection.– : no detection signal.Figure 9. Efficient DNA preparation with the DNeasy mericon Food Kit. Examples of difficult food types were prepared for analysis using the DNeasymericon Food Kit. Cornflakes and ketchup are highly processed foods that contain very fragmented DNA, and ketchup is additionally very acidic due to its vinegar content. Chocolate contains several strong PCR inhibitors. The amplification plots show successful detection of plant DNA by amplification of the chloroplast tRNA gene trnL. Each curve is a replicate of the sample preparation. Performed with pure DNA, this assay gives a positive shift in threshold cycle value (CT) of 2.9 – 3.5 when a sample is diluted 1:10. The shift in threshold cycle value CT between the undiluted and diluted samples fell in this range, demonstrating very efficient inhibitor removal during DNA preparation.mericon PCR Assays offer a highly standardized allergen testing method that enables truecomparison of results across different food types, allergen types, and testing laboratories. One reason for this standardization is the binding specificity of the PCR primers and probes included in the kits. A second reason is the resistance of these assays to the effects of inhibitors present in food matrices. A third reason is that, unlike protein-based assays, DNA detection is not susceptible to molecular modification caused by food processing. This is in contrast to the variability in results from ELISA tests due to the nature of antibody design and the dependence of antibody binding on reaction conditions and protein configuration.Coupled to the high performance of mericon PCR Assays is the efficient DNA extraction delivered by the DNeasy mericon Food Kit. Designed to cope with a spectrum of food types, this sample preparation kit enables high-quality DNA extraction from even highly processed foods where allergen DNA content is low (Figure 9).Allergens¦n Detection of allergen DNA is more reliable then testing protein panels because not all allergenic proteins in food are known and are consequently not included in ELISA panels.¦n DNA extraction is highly standardized and yields purified DNA, whereas protein extraction is typically not specific for target or food matrix.¦n DNA is detectable even in highly processed foods, while proteins are easily denatured or react with components of the food matrix and therefore become undetectable.¦n In contrast to ELISA, PCR enables differentiation of closely-related species (e.g., celery and other umbelliferous plants, or tree nuts).¦n PCR assays include a number of process controls that enhance overall process safety.Furthermore, detection of allergen DNA offers additional advantages:Food Safety Testing 10/2010 Sample & Assay Technologies 110246810125 10 15 20 25 30 35 40Normalized fluorescenceUndilutedDiluted 1:10CycleCornflakes0246810125 10 15 20 25 30 35 40UndilutedDiluted 1:1014CycleChocolate024681012145 10 15 20 25 30 35 40UndilutedDiluted 1:10CycleKetchupIngredient authenticityThe identity of ingredients included in processed food, animal feed, and pharmaceutical products is of growing importance for food safety testing. Materials used in the production of food originate from an increasingly diverse and international list of suppliers and thus, ascertaining the identity of plant and animal matter included in a food ensures maintenance of standards that address food fraud (the use of low-quality matter in high-end foods), cultural specifications (e.g., Kosher and Halal food), and health risks associated with the consumption of undisclosed ingredients.Traditionally, assessment of the plant or animal origin of materials included in food has been performed with ELISA-based assays and protein profiling using two-dimensional pulsed-field gel electrophoresis (2D PFGE). Both methods detect the presence of proteins specific to a particular species of plant or animal and thus, are susceptible to protein configuration changes (denaturation) that result from food processing. Variability in protein structure decreases antibody binding and impacts molecule migration within a gel. Consequently, results with these methods are not always reliable. Furthermore, both methods are labor intensive and require optimization. In contrast, detection of DNA originating from particular animal or plant species by real-time PCR is not vulnerable to processing modifications and thus, offers a highly reliable and robust alternative testing method. The specificity of mericon PCR Assays isevident in the cross-reactivity results in Table 3. A positive detection signal is obtained for domestic pig and wild boar, but not for other animals tested.12 www.qiagen.com Food Safety Testing 10/2010Detection that is robust to food processingSpecies ResultPig +Wild boar +Chicken –Turkey –Sheep –Goat –Cattle –Goose –Horse –Deer –Ostrich –Kangaroo –Moose –Cod –Salmon –Duck –Reindeer (Caribou) –Table 3. Results from cross-reactivity experiments for the mericon Pig Kit + : positive detection.– : no detection signal.Ingredient authenticitySensitivity to deal with challenging starting materialsAnimal and plant matter are often used as subtle food additives. Due to small amounts and the often highly processed nature of the food containing the additive, detecting the presence of animal or plant matter can be difficult. Important to reliable detection in these samples is an extraction method that extracts DNA even from difficult food matrices (e.g., meat samples) and an assay with the sensitivity to detect even low levels of adulterant (Figure 10).Figure 10. Reliable detection of trace amounts of pig. The mericon Pig Kit was used to test for pig DNA in a series of diluted samples from high to very low DNA content. High amounts of DNA do not interfere with the PCR and, conversely, the assay is sensitive enough to detect less than 10 copies of target DNA. References: 1. Pederson, M.H., et al. (2008) Soybean allergen detection methods — a comparison study. Mol. Nutr. Food Res. 52, 1486. Food Safety Testing 10/2010 Sample & Assay Technologies 13Products and expertise to meet your needsQIAGEN’s food testing portfolio is one of many versatile suites of products that QIAGEN brings to your benchtop to facilitate the fundamental work that leads to improvements in life. With this portfolio, we offer our expertise in sample and assay technologies by covering all segments of food safety testing — from pathogens to GMOs, allergens, and plant or animal identification — by streamlining sample preparation and detection protocols, by emphasizing modern methods and technologies, and by focusing on making all of the above accessible and easy to use. 051015202530354045505 10 15 20 25 35 3530Normalized fluorescenceCycle3.1 copies31 copies313 copies3,125 copies31,250 copiesOrdering InformationOrdering InformationProduct Contents Cat. no.mericon Salmonella spp Kit (24)* mericon L. monocytogenes Kit (24)* mericon Campylobacter spp Kit (24)* mericon Campylobacter triple Kit (24)* mericon VTEC stx1/2 Kit (24)* mericon C. sakazakii Kit (24)* mericon S. aureus Kit (24)* mericon Legionella spp Kit (24)* mericon L. pneumophila Kit (24)* mericon Shigella spp Kit (24)* mericon Y. enterocolitica Kit (24)* mericon Screen 35S Kit (24)*PCR Assay Salmonella spp, Positive Control DNA, QuantiTect® Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay L. monocytogenes, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Campylobacter spp, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Campylobacter triple, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay VTEC stx1/2, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay C. sakazakii, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay S. aureus, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Legionella ssp, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay L. pneumophila, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Shigella spp, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Y. enterocolitica, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Screen 35S, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution290013 290023 290033 290043 290053 290063 290073 290083 290093 290103 290113 291013 * Larger kit sizes available; please inquire.14 www.qiagen.com Food Safety Testing 10/2010Discover the mericon food testing portfolio and learn how real-time PCR can revolutionize your testing program Visit www.qiagen.com/mericon-food-safetyOrdering InformationProduct Contents Cat. no.mericon Screen Nos Kit (24)* mericon RR Soy Kit (24)* mericon Pig Kit (24)* mericon Soy Kit (24)* DNeasy mericon Food Kit (50) mericon DNA Bacteria Kit (100) mericon DNA Bacteria Plus Kit (50)PCR Assay Screen Nos, Positive Control Screen DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay RR Soy, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Pig, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution PCR Assay Soy, Positive Control DNA, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, Multiplex PCR Master Mix, 50 x ROX Dye Solution 50 QIAquick® Spin Columns, Proteinase K, buffers Fast Lysis Buffer 50 Pathogen Lysis Tubes L, Fast Lysis Buffer291043 291113 292013 293013 69514 69525 69534* Larger kit sizes available; please inquire.For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at www.qiagen.com or can be requested from QIAGEN Technical Services or your local distributor.Food Safety Testing 10/2010 Sample & Assay Technologies 15Ordering InformationTrademarks: QIAGEN®, QIAquick®, DNeasy ®, mericon™, Q-bond ®, QuantiTect ® (QIAGEN Group); Rotor-Gene® (Corbett Research Pty Ltd); TaqMan® (Roche Group).© 2010 QIAGEN, all rights reserved.1066210 10/2010www.qiagen.comAustralia n Orders 1-800-243-800 n Fax 03-9840-9888 n Technical 1-800-243-066Austria n Orders 0800-28-10-10 n Fax 0800-28-10-19 n Technical 0800-28-10-11Belgium n Orders 0800-79612 n Fax 0800-79611 n Technical 0800-79556Brazil n Orders 0800-557779 n Fax 55-11-5079-4001 n Technical 0800-557779Canada n Orders 800-572-9613 n Fax 800-713-5951 n Technical 800-DNA-PREP (800-362-7737)China n Orders 86-21-3865-3865 n Fax 86-21-3865-3965 n Technical 800-988-0325Denmark n Orders 80-885945 n Fax 80-885944 n Technical 80-885942Finland n Orders 0800-914416 n Fax 0800-914415 n Technical 0800-914413France n Orders 01-60-920-920 n Fax 01-60-920-925 n Technical 01-60-920-930Germany n Orders 02103-29-12000 n Fax 02103-29-22000 n Technical 02103-29-12400Hong Kong n Orders 800 933 965 n Fax 800 930 439 n Technical 800 930 425Ireland n Orders 1800 555 049 n Fax 1800 555 048 n Technical 1800 555 061Italy n Orders 800-789 544 n Fax 02-334304-826 n Technical 800-787980Japan n Telephone 03-6890-7300 n Fax 03-5547-0818 n Technical 03-6890-7300Korea (South) n Orders 080-000-7146 n Fax 02-2626-5703 n Technical 080-000-7145Luxembourg n Orders 8002-2076 n Fax 8002-2073 n Technical 8002-2067Mexico n Orders 01-800-7742-639 n Fax 01-800-1122-330 n Technical 01-800-7742-436The Netherlands n Orders 0800-0229592 n Fax 0800-0229593 n Technical 0800-0229602Norway n Orders 800-18859 n Fax 800-18817 n Technical 800-18712Singapore n Orders 1800-742-4362 n Fax 65-6854-8184 n Technical 1800-742-4368 Spain n Orders 91-630-7050 n Fax 91-630-5145 n Technical 91-630-7050Sweden n Orders 020-790282 n Fax 020-790582 n Technical 020-798328Switzerland n Orders 055-254-22-11 n Fax 055-254-22-13 n Technical 055-254-22-12UK n Orders 01293-422-911 n Fax 01293-422-922 n Technical 01293-422-999USA n Orders 800-426-8157 n Fax 800-718-2056 n Technical 800-DNA-PREP (800-362-7737)Sample & Assay Technologies