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.

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.

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.

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.

VIDAS® Listeria species Xpress (LSX)

2013 ◽  
Vol 96 (2) ◽  
pp. 242-245 ◽  
Author(s):  
Ronald Johnson ◽  
John Mills
Keyword(s):  
Stainless Steel ◽  
Reference Method ◽  
Culture Method ◽  
Culture Methods ◽  
Chi Square ◽  
Listeria Species ◽  
Standard Culture ◽  
Inspection Service ◽  
Chi Square Analysis ◽  
Positive Results

Abstract The AOAC GovVal study compared the VIDAS®Listeria species Xpress (LSX) to the Health Products and Food Branch MFHPB-30 reference method for detection of Listeria on stainless steel. The LSX method utilizes a novel and proprietary enrichment media, Listeria Xpress broth, enabling detection of Listeria species in environmental samples with the automated VIDAS in a minimum of 26 h. The LSX method also includes the use of the chromogenic media, chromID™ Ottaviani Agosti Agar (OAA) and chromID™ Lmono for confirmation of LSX presumptive results. In previous AOAC validation studies comparing VIDAS LSX to the U. S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) and the U. S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) reference methods, the LSX method was approved as AOAC Official Method2010.02 for the detection of Listeria species in dairy products, vegetables, seafood, raw meats and poultry, and processed meats and poultry, and as AOAC Performance Tested Method 100501 in a variety of foods and on environmental surfaces. The GovVal comparative study included 20 replicate test portions each at two contamination levels for stainless steel where fractionally positive results (5–15 positive results/20 replicate portions tested) were obtained by at least one method at one level. Five uncontaminated controls were included. In the stainless steel artificially contaminated surface study, there were 25 confirmed positives by the VIDAS LSX assay and 22 confirmed positives by the standard culture methods. Chi-square analysis indicated no statistical differences between the VIDAS LSX method and the MFHPB-30 standard methods at the 5% level of significance. Confirmation of presumptive LSX results with the chromogenic OAA and Lmono media was shown to be equivalent to the appropriate reference method agars. The data in this study demonstrate that the VIDAS LSX method is an acceptable alternative method to the MFHPB-30 standard culture method for the detection of Listeria species on stainless steel.

Evaluation of the Thermo Scientific™ SureTect™ Listeria species Assay

2014 ◽  
Vol 97 (2) ◽  
pp. 521-538 ◽  
Author(s):  
Jonathan Cloke ◽  
Katharine Evans ◽  
David Crabtree ◽  
Annette Hughes ◽  
Helen Simpson ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Reference Method ◽  
Probability Of Detection ◽  
Stainless Steel Surface ◽  
Specific Method ◽  
Listeria Species ◽  
Smoked Salmon ◽  
Significant Difference ◽  
Thermo Fisher Scientific ◽  
High Level

Abstract The Thermo Scientific™ SureTect™ Listeria species Assay is a new real-time PCR assay for the detection of all species of Listeria in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested MethodsSM program to validate the SureTect Listeria species Assay in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996 including amendment 1:2004 in a variety of foods plus plastic and stainlesssteel. The food matrixes validated were smoked salmon, processed cheese, fresh bagged spinach, cantaloupe, cooked prawns, cooked sliced turkey meat, cooked sliced ham, salami, pork frankfurters, and raw ground beef. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. Inaddition, three matrixes (pork frankfurters, fresh bagged spinach, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled independent laboratory study by the University of Guelph, Canada. Using probability of detection statistical analysis, a significant difference infavour of the SureTect assay was demonstrated between the SureTect and reference method for high level spiked samples of pork frankfurters, smoked salmon, cooked prawns, stainless steel, and low-spiked samples of salami. For all other matrixes, no significant difference was seen between the two methods during the study. Inclusivity testing was conducted with 68 different isolates of Listeria species, all of which were detected by the SureTect Listeria species Assay. None of the 33 exclusivity isolates were detected by the SureTect Listeria species Assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside of the recommended parameters open to variation, which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.

Comparative Evaluation of Veriflow®Listeria Species to USDA Culture-Based Method for the Detection of Listeria spp. in Food and Environmental Samples

2017 ◽  
Vol 100 (5) ◽  
pp. 1434-1444 ◽  
Author(s):  
Adam C Joelsson ◽  
Shawn P Terkhorn ◽  
Ashley S Brown ◽  
Amrita Puri ◽  
Benjamin J Pascal ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Ceramic Tile ◽  
Pcr Amplification ◽  
Reference Method ◽  
Probability Of Detection ◽  
Complex Data ◽  
Listeria Species ◽  
Environmental Surfaces ◽  
Study Results ◽  
Significant Difference

Abstract Veriflow®Listeria species (Veriflow LS) is a molecular-based assay for the presumptive detection of Listeria spp. from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile) and ready-to-eat (RTE) food matrixes (hot dogs and deli meat). The assay utilizes a PCRdetection method coupled with a rapid, visual, flow-based assay that develops in 3 min post-PCR amplification and requires only a 24 h enrichment for maximum sensitivity. The Veriflow LS system eliminates the need for sample purification, gel electrophoresis, or fluorophore-based detection of target amplification and does not require complex data analysis. This Performance Tested MethodSM validation study demonstrated the ability of the Veriflow LS assayto detect low levels of artificially inoculated Listeria spp. in six distinct environmental and food matrixes. In each unpaired reference comparison study, probability of detection analysis indicated that there was no significant difference between the Veriflow LS method and the U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guide Chapter 8.08 reference method. Fifty-one strains of various Listeria spp. were detected in the inclusivity study, and 35 nonspecific organisms went undetected in the exclusivity study. The study results show that the Veriflow LS is a sensitive, selective, and robust assay for the presumptive detection of Listeria spp. sampled from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile) and RTE food matrixes (hot dogs and delimeat).

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.

scholarly journals Comparison of the Petrifilm Dry Rehydratable Film and Conventional Culture Methods for Enumeration of Yeasts and Molds in Foods: Collaborative Study

1997 ◽  
Vol 80 (4) ◽  
pp. 806-824 ◽  
Author(s):  
Michael T Knight ◽  
Melissa C Newman ◽  
M Joseph Benzinger ◽  
Karen L Neufang ◽  
James R Agin ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Food Type ◽  
Corn Meal ◽  
Culture Methods ◽  
Plate Method ◽  
Conventional Culture ◽  
Significant Difference ◽  
Yeasts And Molds ◽  
High Level ◽  
Mold Count

Abstract A collaborative study was performed involving 18 laboratories and 6 food types to compare 3M Petrifilm yeast and mold count plates with the method described in the U.S. Food and Drug Administration’s Bacteriological Analytical Manual. Four species of mold and 2 species of yeast were used to inoculate the following foods: hot dogs, corn meal, ketchup, orange juice, yogurt, and cake mix. Each collaborator received 15 samples of each food type: 5 low-level inoculations, 5 high- level inoculations, and 5 uninoculated samples. There was no significant difference between the means of the 2 methods for any product or inoculation level. The Petrifilm yeast and mold count plate method for enumeration of yeasts and molds in foods has been adopted first action by AOAC INTERNATIONAL.

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