iQ-Check Listeria spp. Real-Time PCR Test Kit

2013 ◽  
Vol 96 (3) ◽  
pp. 508-515
Author(s):  
Wendy F Lauer ◽  
Jean-Philippe Tourniaire
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Reference Method ◽  
Sampling Area ◽  
Fermented Sausage ◽  
Low Level ◽  
Test Kit ◽  
Significant Difference ◽  
High Level ◽  
Health Canada

Abstract A comparative evaluation study of the Bio-Rad® iQ-Check™Listeria species Kit (Bio-Rad Laboratories, Hercules, CA) was conducted at Q Laboratories, Inc., Cincinnati, OH. iQ-Check is a rapid method based on real-time PCR amplification and detection of all species of Listeria, including L. grayi, in food and environmental samples. The iQ-Check method was compared to the Health Canada MFHPB-30 reference method for the analysis of five ready-to-eat meats—deli turkey, hot dogs, liver paté, raw fermented sausage, and deli ham—and one stainless steel surface. Each food matrix was analyzed at two contamination levels: a low level at 0.2–2 CFU/25 g and a high level at 2–5 CFU/25 g. The environmental surfaces were analyzed at a low level of 0.2–2 CFU/5 cm2 sampling area and a high level of 2–5 CFU/5 cm2 sampling area. There were 20 replicates per contamination level and five control replicates at 0 CFU/25 g or 0 CFU/5 cm2 sampling area (uninoculated). All samples that were detected by iQ-Check were subsequently confirmed by reference method protocol. There was no significant difference in the number of positive samples detected by the iQ-Check Listeria spp. Kit in comparison to the Health Canada MFHPB-30 method for all matrixes tested.

iQ-Check Listeria monocytogenes II

2013 ◽  
Vol 96 (3) ◽  
pp. 516-523
Author(s):  
Wendy F Lauer ◽  
Jean-Philippe Tourniaire
Keyword(s):  
Listeria Monocytogenes ◽  
Reference Method ◽  
Contamination Level ◽  
Stainless Steel Surface ◽  
Sampling Area ◽  
Fermented Sausage ◽  
Low Level ◽  
Significant Difference ◽  
High Level ◽  
Health Canada

Abstract A comparative evaluation study of the Bio-Rad® iQ-Check™Listeria monocytogenes II Kit (Bio-Rad Laboratories, Hercules, CA) was conducted at Q Laboratories, Inc., Cincinnati, OH. iQ-Check is a rapid method based on real-time PCR amplification and detection of L. monocytogenes in food and environmental samples. The iQ-Check method was compared to the Health Canada MFHPB-30 reference method for the analysis of five ready-to-eat meats—deli turkey, hot dogs, liver paté, raw fermented sausage, and deli ham—and one stainless steel surface. Each food matrix was analyzed at two contamination levels: a low level at 0.2–2 CFU/25 g and a high level at 2–5 CFU/25 g. The environmental surfaces were analyzed at a low level of 0.2–2 CFU/5 cm2 sampling area and a high level of 2–5 CFU/5 cm2 sampling area. There were 20 replicates per contamination level and five control replicates at 0 CFU/25 g or 0 CFU/5 cm2 sampling area (uninoculated). All samples detected by iQ-Check were subsequently confirmed by reference method protocol. There was no significant difference in the number of positive samples detected by the iQ-Check Listeria monocytogenes II Kit in comparison to the Health Canada MFHPB-30 method for all matrixes tested.

3M™ Tecra™ Listeria Visual Immunoassay: AOAC Official MethodsSM 995.22 and 2002.09

2013 ◽  
Vol 96 (2) ◽  
pp. 218-224
Author(s):  
DeAnn L Benesh ◽  
Erin S Crowley ◽  
Patrick M Bird
Keyword(s):  
Stainless Steel ◽  
Reference Method ◽  
Sampling Area ◽  
Inoculum Level ◽  
Fermented Sausage ◽  
High Inoculum ◽  
Significant Difference ◽  
Environmental Surface ◽  
High Level ◽  
Health Canada

Abstract A validation study of the 3M™ Tecra™ Listeria Visual Immunoassay (VIA; 3M Food Safety, St. Paul, MN) was conducted at Q Laboratories, Inc., Cincinnati, OH. The 3M Tecra Listeria VIA method was compared to the Health Canada MFHPB-30 reference method for the analysis of five ready-to-eat (RTE) meats: deli turkey, hot dogs, liver pate, raw fermented sausage, and deli ham, and on a stainless steel environmental surface. Twenty replicates of each of the five food matrixes were analyzed at a low and a high inoculum level. The low-level test portions were inoculated with 0.2–2 CFU/25 g, and the high-level test portions with 2–5 CFU/25 g. In addition, 20 replicates of one environmental surface were analyzed at a low and a high inoculum level. The low-level sampling area was inoculated with 0.2–2 CFU/5 cm2, and the high-level area with 2–5 CFU/5 cm2. Five control replicates were also analyzed at 0 CFU/25 g (uninoculated) for the foods and at 0 CFU/5 cm2 for the environmental sampling area. There was no significant difference in the number of positives detected by the 3M Tecra Listeria VIA and the Health Canada MFHPB-30 reference method for four of the RTE meats and the stainless steel environmental surface analyzed in this study. For the raw, fermented sausage, there was a significant difference in the number of positives detected for the high inoculum level by the 3M Tecra Listeria VIA and the Health Canada MFHPB-30 reference method, with the 3M Tecra Listeria VIA method detecting more positives.

scholarly journals Validation of iQ-Check E. coli O157:H7 Real-Time PCR Test Kit for Detection of Escherichia coli O157:H7 in Selected Foods

2009 ◽  
Vol 92 (4) ◽  
pp. 1095-1104 ◽  
Author(s):  
Wendy F Lauer ◽  
Sylvie Tymciu ◽  
Caroline D Sidi ◽  
Pierre Sonigo
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Ground Beef ◽  
Reference Method ◽  
Target Sequence ◽  
Apple Cider ◽  
E Coli ◽  
Test Kit ◽  
Significant Difference ◽  
The U.S

Abstract iQ-Check E. coli O157:H7 (Bio-Rad Laboratories, Hercules, CA) is a real-time PCR kit for detection of E. coli O157:H7 from selected foods. Specific fluorescent oligonucleotide probes are used to detect target DNA during the amplification, by hybridizing to the amplicons. These fluorescent probes are linked to a fluorophore which fluoresces only when hybridized to the target sequence. Three foods (ground beef, apple cider, fresh spinach) were selected to compare the performance of iQ-Check E. coli O157:H7 to the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG) reference method for ground beef and the U.S. Food and Drug Administration/Bacteriological Analytical Manual reference method for apple cider and fresh spinach. Three protocols were tested in this study: a shortened 8 h primary enrichment in buffered peptone water (BPW), a 24 h enrichment in BPW, and an enrichment in appropriate reference method enrichment broth. The iQ-Check E. coli O157:H7 method was able to identify more true/confirmed positive samples than the reference method. Inclusivity and exclusivity rates of the method were 100. iQ-Check E. coli O157:H7 performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no significant difference observed in performance over the shelf life of the kit.

3M™ Petrifilm™ Environmental Listeria Plate

2013 ◽  
Vol 96 (2) ◽  
pp. 225-228 ◽  
Author(s):  
DeAnn L Benesh ◽  
Erin S Crowley ◽  
Patrick M Bird
Keyword(s):  
Stainless Steel ◽  
Reference Method ◽  
Sampling Area ◽  
Plate Method ◽  
Inoculum Level ◽  
Environmental Surfaces ◽  
Significant Difference ◽  
Environmental Surface ◽  
High Level ◽  
Health Canada

Abstract A validation study of the 3M™ Petrifilm™ Environmental Listeria (EL) Plate (3M Food Safety, St. Paul, MN) was conducted at Q Laboratories, Inc., Cincinnati, OH. The method was compared to the Health Canada MFHPB-30 reference method for the analysis of stainless steel environmental surfaces. Twenty replicates of the environmental surface were analyzed at a low and a high inoculum level. The low-level sampling area was inoculated with 0.2–2 CFU/5 cm2, and the high-level sampling area was inoculated with 2–5 CFU/5 cm2. Five control replicates were also analyzed at 0 CFU/5 cm2. There was no significant difference in the number of positives detected by the 3M Petrifilm EL Plate method and the Health Canada MFHPB-30 reference method for the environmental surface analyzed in this study.

MicroSEQ®Listeria spp. Detection Kit

2013 ◽  
Vol 96 (3) ◽  
pp. 532-541
Author(s):  
Olga Petrauskene ◽  
Yan Cao ◽  
Patrick Zoder ◽  
Lily Y Wong ◽  
Priya Balachandran ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Reference Method ◽  
Sampling Area ◽  
Culture Methods ◽  
Listeria Species ◽  
Significant Difference ◽  
Environmental Surface ◽  
High Level ◽  
Health Canada ◽  
Positive Results

Abstract The Applied Biosystems Performance Tested MethodSM for detecting Listeria species in food and environmental samples was compared to the Health Canada reference method (MFHPB-30) for the analysis of five ready-to-eat (RTE) meats (deli turkey, hot dogs, liver paté, deli ham, and raw fermented sausage) and a stainless steel surface. The MicroSEQ method includes the MicroSEQ®Listeria spp. Detection Kit and the option of two different sample preparation kits, either the automated high-throughput PrepSEQ™ Nucleic Acid Extraction Kit or the manual low- to mid-throughput PrepSEQ™ Rapid Spin Sample Preparation Kit. For each sample matrix, 20 replicates were analyzed at two inoculum levels: for RTE meats a low-level inoculum at 0.2–2 CFU/25 g and a high-level inoculum at 2–5 CFU/25 g; and for environmental surfaces, a low-level inoculum at 0.2–2 CFU/5 cm2 sampling area and a high-level inoculum at 2–5 CFU/5 cm2 sampling area. Five control replicates were also analyzed at 0 CFU/25 g (uninoculated) for food or 0 CFU/5 cm2 sampling area for environmental surface. Both sample preparation methods returned identical results. There was no statistically significant difference in the number of positive samples detected by the MicroSEQ Listeria species method and the MFHPB-30 reference method for three RTE meats and for the one stainless steel environmental surface tested. For deli turkey, there was a statistically significant difference in the number of positive results detected by the MicroSEQ method and the Health Canada MFHPB-30 reference method for the low inoculation level, with the MicroSEQ method detecting more positives. For hot dogs, statistical equivalence was not applicable since hot dogs were spiked with 10x Listeria innocua as competitive background, and the MicroSEQ method detects all known Listeria. Because the MicroSEQ method uses real-time PCR to detect pathogens, it provides faster time-to-results with equivalent detection compared to culture methods. The MicroSEQ method detects Listeria species within 2 to 3 h following 24 to 28 h enrichment compared to culture methods that take at least 5 days for presumptive positive results.

MicroSEQ®Listeria monocytogenes Detection Kit

2013 ◽  
Vol 96 (3) ◽  
pp. 542-551
Author(s):  
Robert S Tebbs ◽  
Yan Cao ◽  
Priya Balachandran ◽  
Lily Y Wong ◽  
Olga Petrauskene
Keyword(s):  
Listeria Monocytogenes ◽  
Sample Preparation ◽  
Reference Method ◽  
Sampling Area ◽  
Culture Methods ◽  
Significant Difference ◽  
Environmental Surface ◽  
High Level ◽  
Health Canada ◽  
Positive Results

Abstract The Applied Biosystems Performance Tested MethodSM for detecting Listeria monocytogenes in food and environmental samples was compared to the Health Canada reference method (MFHPB-30) for the analysis of five ready-to-eat (RTE) meats (deli turkey, hot dogs, liver paté, deli ham, and raw fermented sausage) and a stainless steel environmental surface. The MicroSEQ® method includes the MicroSEQListeria monocytogenes Detection Kit and the option of two different sample preparation kits, either the automated high-throughput PrepSEQ™ Nucleic Acid Extraction Kit or the manual low- to mid-throughput PrepSEQ Rapid Spin Sample Preparation Kit. For each sample matrix, 20 replicates were analyzed at two inoculum levels: for RTE meats, a low-level inoculum at 0.2–2 CFU/25 g and a high-level inoculum at 2–5 CFU/25 g; and for environmental surfaces, a low-level inoculum at 0.2–2 CFU/5 cm2 sampling area and a high-level inoculum at 2–5 CFU/5 cm2 sampling area. Five control replicates were also analyzed at 0 CFU/25 g (uninoculated) for food or 0 CFU/5 cm2 sampling area for environmental surface. Both sample preparation methods returned identical results. There was no statistically significant difference in the number of positive samples detected by the MicroSEQ Listeria monocytogenes method and the MFHPB-30 reference method for four of the five RTE meats and the one stainless steel surface tested. For deli turkey, there was a statistically significant difference in the number of positive results detected by the MicroSEQ method and the Health Canada MFHPB-30 reference method for the low inoculation level, with the MicroSEQ method detecting more positives. Because the MicroSEQ method uses real-time PCR to detect pathogens, it provides faster time-to-results compared to culture methods while at the same time demonstrating equivalent detection. The MicroSEQ method detects L. monocytogenes within 2 to 3 h following 24 to 28 h enrichment compared to culture methods that take at least 5 days for presumptive positive results.

scholarly journals Evaluation of Applied Biosystems MicroSEQ® Real-Time PCR System for Detection of Salmonella spp. in Food

2011 ◽  
Vol 94 (4) ◽  
pp. 1106-1116 ◽  
Author(s):  
Priya Balachandran ◽  
Yanxiang Cao ◽  
Lily Wong ◽  
Manohar R Furtado ◽  
Olga V Petrauskene ◽  
...  
Keyword(s):  
Real Time ◽  
Study Design ◽  
Real Time Pcr ◽  
Detection System ◽  
Peanut Butter ◽  
Reference Method ◽  
Black Pepper ◽  
Salmonella Spp ◽  
Culture Methods ◽  
Significant Difference

Abstract Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time-to-results compared to traditional culture methods. In this study, the MicroSEQ® real-time PCR system was evaluated for detection of Salmonella spp. in 10 different food matrixes following the AOAC Research Institute's Performance Tested MethodSM validation program. In addition, the performance of the MicroSEQ system was evaluated for the detection of Salmonella in peanut butter as a part of the Emergency Response Validation Program sponsored by the AOAC Research Institute. The system was compared to the ISO 6579 reference method using a paired-study design for detecting Salmonella spp. in raw ground beef, raw chicken, raw shrimp, Brie cheese, shell eggs, cantaloupe, chocolate, black pepper, dry infant formula, and dry pet food. For the peanut butter study, the system was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual procedures using an unpaired-study design. No significant difference in performance was observed between the MicroSEQ Salmonella spp. detection system and the corresponding reference methods for all 11 food matrixes. The MicroSEQ system detected all Salmonella strains tested, while showing good discrimination against detection of an exclusivity panel of 30 strains, with high accuracy.

Interlaboratory Validation for a Real-Time PCR Salmonella Detection Method Using the ABI 7500 FAST Real-Time PCR System

2015 ◽  
Vol 78 (6) ◽  
pp. 1119-1124 ◽  
Author(s):  
CHORNG-MING CHENG ◽  
TARA DORAN ◽  
WEN LIN ◽  
KAI-SHUN CHEN ◽  
DONNA WILLIAMS-HILL ◽  
...  
Keyword(s):  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
Detection Method ◽  
Reference Method ◽  
Network Member ◽  
Fast System ◽  
Enhanced Sensitivity ◽  
Significant Difference ◽  
Chili Powder

Sixteen FERN (Food Emergency Response Network) member laboratories collaborated in this study to verify extension of the real-time PCR Salmonella detection method originally designed for the single-tube Cepheid SmartCycler II and validated against the Salmonella method of the U.S. Food and Drug Administration Bacteriological Analytical Manual to the Applied Biosystems (ABI) 7500 FAST Real-Time PCR system multiwell plate platform. Four foods were selected for this study: chili powder, soft cheese, fish, and tomatoes; these foods represent products that are commonly analyzed for the presence of Salmonella for regulatory purposes. Each food consisted of six uninoculated control samples, six samples inoculated with low Salmonella levels (target 1 to 5 CFU/25 g), and six samples inoculated with high levels (target 10 to 50 CFU/25 g). All samples were tested for Salmonella using the 24-h quantitative PCR (qPCR) method for detecting Salmonella, which utilizes modified buffered peptone water as the sole enrichment medium and an internal control for the qPCR. Each of these 18 samples was individually analyzed for Salmonella by the collaborating laboratories using both the ABI 7500 FAST system (alternative method) and the SmartCycler II system (reference method). Statistical analysis of the data revealed no significant difference (P ≥ 0.05) between these two qPCR platforms except for the chili powder samples. The differences noted with chili powder (P = 0.0455) were attributed to the enhanced sensitivity of the ABI 7500 FAST system compared with the SmartCycler II system. The detection limit of both qPCR methods was 0.02 to 0.15 CFU/g. These results provide a solid basis for extending the 24-h qPCR Salmonella method to the ABI 7500 FAST system for high-throughput detection of Salmonella in foods.

scholarly journals iQ-Check Salmonella II: Real-Time Polymerase Chain Reaction Test Kit

2009 ◽  
Vol 92 (6) ◽  
pp. 1865-1870 ◽  
Author(s):  
Wendy F Lauer ◽  
Caroline D Sidi ◽  
Jean-Philippe Tourniaire ◽  
Thomas Hammack
Keyword(s):  
Real Time ◽  
Peanut Butter ◽  
Reference Method ◽  
Polymerase Chain Reaction Test ◽  
Target Sequence ◽  
Chi Square ◽  
Test Kit ◽  
Significant Difference ◽  
Chi Square Analysis ◽  
Chain Reaction Test

Abstract iQ-Check Salmonella II is a real-time PCR kit for detection of Salmonella in foods. Specific oligonucleotide probes are used to detect target DNA during the amplification, by hybridizing to the amplicons. These probes are linked to a fluorophore, which fluoresces only when hybridized to the target sequence. As part of an Emergency Response Validation due to a massive outbreak and subsequent recall, peanut butter was tested to compare the performance of iQ-Check Salmonella II to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) reference method for detection of Salmonella. A single enrichment in buffered peptone water was used for a reduced enrichment time of 21 1 h over the 48 h reference method. There was no significant difference in the performance of the iQ-Check kit when compared to the FDA-BAM method, as determined by Chi-square analysis. All samples identified as positive by iQ-Check were confirmed by reference method protocol.

Detection of Salmonella species in a Variety of Foods by the DuPont™ BAX® System Real-Time PCR Assay for Salmonella: First Action 2013.02

2014 ◽  
Vol 97 (3) ◽  
pp. 868-875 ◽  
Author(s):  
F Morgan Wallace ◽  
Bridget Andaloro ◽  
Dawn Fallon ◽  
Nisha Corrigan ◽  
Stephen Varkey ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Orange Juice ◽  
Collaborative Study ◽  
Reference Method ◽  
Probability Of Detection ◽  
Test Method ◽  
Pcr Assay ◽  
Test Portion ◽  
Significant Difference

Abstract A multilaboratory study was conducted to evaluate the ability of the DuPont™ BAX® System Real-Time PCR Assay for Salmonella to detect the target species in a variety of foods and environmental surfaces. Internal validation studies were performed by DuPont Nutrition & Health on 24 different sample types to demonstrate the reliability of the test method among a wide variety of sample types. Two of these matrixes—pork and turkey frankfurters and pasteurized, not-from-concentrate orange juice without pulp—were each evaluated in 14 independent laboratories as part of the collaborative study to demonstrate repeatability and reproducibility of the internal laboratory results independent of the end user. Frankfurter samples were evaluated against the U. S. Department of Agriculture, Food Safety and Inspection Service reference method as a paired study, while orange juice samples were evaluated against the U. S. Food and Drug Administration reference method as an unpaired study, using a proprietary media for the test method. Samples tested in this study were artificially inoculated with a Salmonella strain at levels expected to produce low (0.2–2.0 CFU/test portion) or high (5 CFU/test portion) spike levels on the day of analysis. For each matrix, the collaborative study failed to show a statistically significant difference between the candidate method and the reference method using the probability of detection statistical model.

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