Evaluation of the Thermo Scientific™ SureTect™ Salmonella species Assay

2014 ◽  
Vol 97 (2) ◽  
pp. 539-560
Author(s):  
Jonathan Cloke ◽  
Dorn Clark, Jr ◽  
Roy Radcliff ◽  
Carlos Leon-Velarde ◽  
Nathan Larson ◽  
...  
Keyword(s):  
Validation Study ◽  
Reference Method ◽  
Probability Of Detection ◽  
Chicken Breast ◽  
Stainless Steel Surface ◽  
Specific Method ◽  
Surface Samples ◽  
Significant Difference ◽  
Liquid Whole Egg ◽  
Whole Egg

Abstract The Thermo Scientific™ SureTect™ Salmonella species Assay is a new real-time PCR assay for the detection of Salmonellae in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested MethodsSM program to validate the SureTect Salmonella species Assay incomparison to the reference method detailed in International Organization for Standardization 6579:2002 in a variety of food matrixes, namely, raw ground beef, raw chicken breast, raw ground pork, fresh bagged lettuce, pork frankfurters, nonfat dried milk powder, cooked peeled shrimp, pasteurized liquid whole egg, ready-to-eat meal containing beef, and stainlesssteel surface samples. With the exception of liquid whole egg and fresh bagged lettuce, which were tested in-house, all matrixes were tested by Marshfield Food Safety, Marshfield, WI, on behalf of Thermo Fisher Scientific. In addition, three matrixes (pork frankfurters, lettuce, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled laboratory study by the University of Guelph, Canada. No significant difference by probability of detection or McNemars Chi-squared statistical analysis was found between the candidate or reference methods for any of the food matrixes or environmental surface samples tested during the validation study. Inclusivity and exclusivity testing was conducted with 117 and 36 isolates, respectively, which demonstrated that the SureTect Salmonella species Assay was able to detect all the major groups of Salmonella enterica subspecies enterica (e.g., Typhimurium) and the less common subspecies of S. enterica (e.g., arizoniae) and the rarely encountered S. bongori. None of the exclusivity isolates analyzed were detected by the SureTect Salmonella 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 (enrichment time and temperature, and lysis temperature), which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.

Validation of the Applied Biosystems 7500 Fast Instrument for Detection of Listeria monocytogenes with the SureTect Listeria monocytogenes PCR Assay

2016 ◽  
Vol 99 (3) ◽  
pp. 676-685
Author(s):  
Jonathan Cloke ◽  
Julia Arizanova ◽  
David Crabtree ◽  
Helen Simpson ◽  
Katharine Evans ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Reference Method ◽  
Probability Of Detection ◽  
International Organization ◽  
Stainless Steel Surface ◽  
Pcr Assay ◽  
Surface Samples ◽  
Wide Range ◽  
Significant Difference ◽  
Level 2

Abstract In 2013, the Thermo Scientific™ SureTect™ Listeria monocytogenes Real-Time PCR Assay was certified by the AOAC Research Institute (RI) Performance Tested MethodsSM program as a rapid method for the detection of L. monocytogenes from a wide range of food matrixes and surface samples. This report details the method modification studies undertaken to extend the analysis of this PCR assay to the Applied Biosystems™ 7500 Fast PCR Instrument and Applied Biosystems RapidFinder™ Express 2.0 software allowing the use of the SureTect assay on a 96 well format PCR cycler in addition to the current workflow, which uses the 24 well Thermo Scientific PikoReal™ PCR Instrument and Thermo Scientific SureTect software. Because this study was deemed by AOAC-RI to be a level 2 method modification study, a representative range of food matrixes covering raw ground turkey, 2% fat pasteurized milk, and bagged lettuce as well as stainless steel surface samples were analyzed with the Applied Biosystems 7500 Fast PCR Instrument and RapidFinder Express 2.0 software. All testing was conducted in comparison to the reference method detailed in International Organization for Standardization (ISO) 6579:2002. No significant difference by probability of detection statistical analysis was found between the SureTect Listeria monocytogenes PCR Assay or the ISO reference method methods for any of the matrixes analyzed during the study.

Validation of the Applied Biosystems 7500 Fast Instrument for Detection of Listeria Species with the SureTect Listeria Species PCR Assay

2016 ◽  
Vol 99 (2) ◽  
pp. 407-416
Author(s):  
Jonathan Cloke ◽  
Julia Arizanova ◽  
David Crabtree ◽  
Helen Simpson ◽  
Katharine Evans ◽  
...  
Keyword(s):  
Reference Method ◽  
Probability Of Detection ◽  
Stainless Steel Surface ◽  
Pcr Assay ◽  
Listeria Species ◽  
Surface Samples ◽  
Wide Range ◽  
Significant Difference ◽  
The Matrix ◽  
Level 2

Abstract The Thermo Scientific™ SureTect™ Listeria species Real-Time PCR Assay was certified during 2013 by the AOAC Research Institute (RI) Performance Tested MethodsSM program as a rapid method for the detection of Listeria species from a wide range of food matrixes and surface samples. A method modification study was conducted in 2015 to extend the matrix claims of the product to a wider range of food matrixes. This report details the method modification study undertaken to extend the use of this PCR kit to the Applied Biosystems™ 7500 Fast PCR Instrument and Applied Biosystems RapidFinder™ Express 2.0 software allowing use of the assay on a 96-well format PCR cycler in addition to the current workflow, using the 24-well Thermo Scientific PikoReal™ PCR Instrument and Thermo Scientific SureTect software. The method modification study presented in this report was assessed by the AOAC-RI as being a level 2 method modification study, necessitating a method developer study on a representative range of food matrixes covering raw ground turkey, 2% fat pasteurized milk, and bagged lettuce as well as stainless steel surface samples. All testing was conducted in comparison to the reference method detailed in International Organization for Standardization (ISO) 6579:2002. No significant difference by probability of detection statistical analysis was found between the SureTect Listeria species PCR Assay or the ISO reference method methods for any of the three food matrixes and the surface samples analyzed during the study.

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.

Evaluation of the Thermo Scientific™ SureTect™ Listeria monocytogenes Assay

2014 ◽  
Vol 97 (1) ◽  
pp. 133-154 ◽  
Author(s):  
Jonathan Cloke ◽  
Carlos Leon-Velarde ◽  
Nathan Larson ◽  
Keron Dave ◽  
Katharine Evans ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Reference Method ◽  
Ice Cream ◽  
Probability Of Detection ◽  
Specific Method ◽  
Food Contact Surfaces ◽  
Significant Difference ◽  
Thermo Fisher Scientific ◽  
Reliable Performance

Abstract The Thermo Scientific™ SureTect™Listeria monocytogenes Assay is a new real-time PCR assay for the detection of Listeria monocytogenes in food and environmental samples. This assay was validated using the AOAC Research Institute (AOAC-RI) Performance Tested MethodsSM program in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996, including Amendment 1:2004 with the following foods and food contact surfaces: smoked salmon, processed cheese, fresh bagged spinach, fresh cantaloupe, cooked prawns (chilled product), cooked sliced turkey meat (chilled product), ice cream, pork frankfurters, salami, ground raw beef meat (12% fat), plastic, and stainless steel. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. In addition, three matrixes (pork frankfurters, bagged lettuce, and stainless steel) were analyzed independently as part of the AOAC-RI controlled laboratorystudy by the University of Guelph, Canada. Using probability of detection (POD) statistical analysis, a significant difference was demonstrated between the candidate and reference methods for salami, cooked sliced turkey and ice cream in favor of the SureTect assay. For all other matrixes, no significant difference by POD was seen between the two methods during the study. Inclusivity and exclusivity testing was also conducted with 53 and30 isolates, respectively, which demonstrated that the SureTect assay was able to detect all serotypes of L. monocytogenes. None of the exclusivity isolates analyzed were detectedby the SureTect assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside the recommended parameters open to variation, i.e., enrichment time and temperature and lysis temperature, which demonstrated that the assay gave reliable performance. Accelerated stability testing was alsoconducted, validating the assay shelf life.

scholarly journals Validation of the One Broth One Plate for Salmonella Method for Detection of Salmonella Spp. in Select Food and Environmental Samples: AOAC Performance Tested MethodSM 102002

2020 ◽  
Author(s):  
Susan Alles ◽  
Brooke Roman ◽  
Quynh-Nhi Le ◽  
Magdalena Kurteu ◽  
Ezzeddine Elmerhebi ◽  
...  
Keyword(s):  
Validation Study ◽  
Environmental Samples ◽  
Reference Method ◽  
Black Pepper ◽  
Single Step ◽  
Probability Of Detection ◽  
Stainless Steel Surface ◽  
Salmonella Spp ◽  
Simple Method ◽  
The U.S

Abstract Background One Broth One Plate for Salmonella (OBOP Salmonella) is a rapid and simple method for detection of Salmonella spp. in food and environmental samples using traditional culture methodology. The method utilizes single-step enrichment followed by plating to a selective/differential, chromogenic agar. Objective The purpose of the validation study was to measure the effectiveness of the OBOP Salmonella method in comparison to reference culture procedures. Methods Performance of the OBOP Salmonella method was compared to that of the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 5 reference method for queso fresco, smoked salmon, cantaloupe, chocolate, black pepper, chili powder, dry pet food, and sponge samples from a stainless steel surface, or to that of the U.S. Department of Agriculture Microbiology Laboratory Guidebook Chapter 4.10 method for raw ground turkey, chicken carcass rinse, and pasteurized liquid egg. Inclusivity/exclusivity, robustness, and stability/lot-to-lot consistency testing was also performed. Results In the matrix study, there were no statistically significant differences in performance between the OBOP Salmonella and reference methods, as determined by probability of detection analysis (P < 0.05), for any of the matrixes examined. All 104 Salmonella spp. strains produced positive results in inclusivity testing, and all 33 non-salmonellae exclusivity strains tested negative with the OBOP Salmonella method. Conclusions Results of the validation study show that the OBOP Salmonella method is a reliable procedure for detection of Salmonella spp. in select matrixes. The method is simple to perform, requires no specialized equipment, and produces results in as little as 37 h.

Evaluation of 3M Molecular Detection Assay (MDA) 2–Listeria for the Detection of Listeria Species in Select Foods and Environmental Surfaces: Collaborative Study, First Action 2016.07

2017 ◽  
Vol 100 (1) ◽  
pp. 82-98
Author(s):  
Patrick Bird ◽  
Jonathan Flannery ◽  
Erin Crowley ◽  
James Agin ◽  
David Goins ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Collaborative Study ◽  
Reference Method ◽  
Probability Of Detection ◽  
Chicken Breast ◽  
Test Portion ◽  
Listeria Species ◽  
Environmental Surfaces ◽  
Detection Assay ◽  
Significant Difference

Abstract 3M Molecular Detection Assay (MDA) 2–Listeria uses loop-mediated isothermal amplification and bioluminescence detection to rapidly detect Listeria species in a broad range of food types and environmental surfaces. Using an unpaired study design, MDA 2–Listeria was compared with the U.S. Department of Agriculture, Food Safety and Inspection Service's Microbiology Laboratory Guidebook Chapter 8.09 “Isolation and identification of Listeria monocytogenes from red meat, poultry and egg products, and environmental samples” reference method for the detection of Listeria in deli turkey and raw chicken breast fillet. Technicians from 13 laboratories located within the continental United States and Canada participated in the collaborative study. Each matrix was evaluated at three levels of contamination: uninoculated control (0 CFU/test portion), low inoculum (0.2–2 CFU/test portion), and high inoculum (2–5 CFU/test portion). Statistical analysis was conducted according to the probability of detection (POD) statistical model. Results obtained for the low-inoculum-level test portions produced a difference between two laboratory POD values (dLPOD) with 95% confidence intervals of 0.04 (–0.08, 0.17) for deli turkey, indicating the difference between the methods was not statistically significant at the P = 0.05. For raw chicken breast fillet, a dLPOD value with 95% confidence interval of 0.16 (0.04, 0.28) indicated a statistically significant difference between the two methods, with an observed higher proportion of positive results by the candidate method than the reference method.

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).

Evaluation of the 3M™ Molecular Detection Assay (MDA) 2 – Listeria monocytogenes for the Detection of Listeria monocytogenes in a Variety of Foods and Select Environmental Surfaces: Collaborative Study, First Action 2016.08

2017 ◽  
Vol 100 (2) ◽  
pp. 454-469
Author(s):  
Patrick Bird ◽  
Jonathan Flannery ◽  
Erin Crowley ◽  
James Agin ◽  
David Goins ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Confidence Interval ◽  
Molecular Detection ◽  
Reference Method ◽  
Probability Of Detection ◽  
Chicken Breast ◽  
Inoculum Level ◽  
Test Portion ◽  
Environmental Surfaces ◽  
Detection Assay

Abstract The 3M™ Molecular Detection Assay (MDA) 2 – Listeria monocytogenes uses loop-mediated isothermal amplification of unique DNA target sequences combined with bioluminescence to rapidly detect Listeria monocytogenes in a broad range of food types and on environmental surfaces. Using an unpaired study design, technicians from 13 laboratories located in the United States and Canada compared the 3M MDA 2 – Listeria monocytogenes to the U.S. Department of Agriculture Food Safety Inspection Service Microbiology Laboratory Guidebook Chapter 8.09 “Isolation and Identification of Listeria monocytogenes from Red Meat, Poultry, and Egg Products, and Environmental Samples” reference method for the detection of L. monocytogenes in deli turkey and raw chicken breast fillet. Each matrix was evaluated at three levels of contamination: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2–2 CFU/test portion), and a high inoculum level (2–5 CFU/test portion). Statistical analysis was conducted according to the probability of detection (POD) statistical model. Results obtained for the low inoculum level test portions produced a difference in the collaborating laboratory POD (dLPOD) value of 0.04 with a 95% confidence interval of (−0.08, 0.17) for deli turkey, indicating that the difference between methods was not statistically significant at the 0.05 probability level. For raw chicken breast fillet, a dLPOD value of 0.16 with a 95% confidence interval of (0.04, 0.28) indicated a statistically significant difference, with an observed higher proportion of positive results by the candidate method compared to the reference method.

Evaluation of VIDAS® UP Salmonella (SPT) Assay for the Detection of Salmonella in a Variety of Foods and Environmental Samples: Collaborative Study

2013 ◽  
Vol 96 (4) ◽  
pp. 808-821 ◽  
Author(s):  
Patrick Bird ◽  
Kiel Fisher ◽  
Megan Boyle ◽  
Travis Huffman ◽  
Marc Juenger ◽  
...  
Keyword(s):  
Confidence Intervals ◽  
Environmental Samples ◽  
Collaborative Study ◽  
Reference Method ◽  
The United States ◽  
Probability Of Detection ◽  
Inoculum Level ◽  
Test Portion ◽  
Low Level ◽  
Significant Difference

Abstract The VIDAS® UP Salmonella (SPT) uses recombinant phage proteins to detect Salmonella species in human and animal food products and production environmental samples after 18–26 h of enrichment. The VIDAS SPT assay is performed with the automated VIDAS or mini-VIDAS instruments. The VIDAS SPT method was compared in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) 4.05 (2011) Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products reference method following the current AOAC guidelines. A total of 15 laboratories representing government, academia, and industry throughout the United States participated. One matrix, raw ground beef, was analyzed using two different test portion sizes, 25 and 375 g. Each test portion was artificially contaminated with Salmonella at three inoculation levels, an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2–2 CFU/test portion), and a high inoculum level (2–5 CFU/test portion). In this study, 1656 unpaired replicate samples were analyzed. Of those unpaired replicates, 476 were presumptive positive by the VIDAS method, with 475 confirmed positive by the traditional confirmation procedures and 476 confirmed positive by an alternative confirmation procedure. There were 411 confirmed positive replicates by the USDA/FSIS-MLG reference method. Statistical analysis was conducted according to the probability of detection (POD). For the low-level 375 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.01 (−0.12, +0.15) for samples confirmed following the traditional confirmation; 0.02 (−0.18, +0.2) for samples confirmed following traditional confirmation on IBISA and ASAP; and 0.03 (−0.18, +0.24) for samples confirmed following the alternative confirmation on IBISA and ASAP. For the low-level 25 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.41, (0.32, +0.49) for samples confirmed following the traditional confirmation, the traditional confirmation on IBISA and ASAP, and the alternative confirmation on IBISA and ASAP. With 0.0 within the confidence intervals for the 375 g test portions, there was no statistically significant difference in the number of positive samples detected by the VIDAS SPT method and the USDA/FSIS-MLG method at the 0.05 level. For the 25 g test portions, a statistically significant difference was observed between the VIDAS SPT method and the reference method for the low inoculum level, where the VIDAS SPT method recovered a higher number of positive results than the reference method. It is recommended that the VIDAS SPT method with the optional ASAP and IBISA agar confirmation method be adopted for Official First Action status for the detection of Salmonella in a variety of foods and environmental samples.

Method Modification of the Thermo Scientific SureTect Listeria monocytogenes Assay for Raw Meat, Dairy, Produce, and Seafood

2015 ◽  
Vol 98 (5) ◽  
pp. 1315-1324 ◽  
Author(s):  
Jonathan Cloke ◽  
Katharine Evans ◽  
David Crabtree ◽  
Annette Hughes ◽  
Helen Simpson ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Reference Method ◽  
Probability Of Detection ◽  
Pcr Assay ◽  
Significant Difference ◽  
Ground Pork ◽  
Thermo Fisher Scientific ◽  
Pork Sausages ◽  
The University ◽  
Fisher Scientific

Abstract The Thermo Scientific™ SureTect™ Listeria monocytogenes assay is a real-time PCR assay for the detection of Listeria monocytogenes in food and environmental samples, which was certified during 2013 by the AOAC Research Institute (RI) as Performance Tested MethodSM (PTM) 061302 for a representative range of key food matrixes and production surfaces. This report details the method modification study, which was conducted during 2014, using the AOAC-RI PTM program to extend the validated matrix claims of the assay in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996, including Amendment 1:2004, to gain certification for raw ground turkey, raw ground pork, pasteurized 2% milk, raw pork sausages, raw cod, pasteurized brie cheese, cooked sliced ham, and bagged lettuce. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. In addition, brie cheese, bagged lettuce, and raw cod were analyzed independently by the University of Guelph, Canada, during the AOAC-RI controlled independent laboratory study. Using probability of detection (POD) statistical analysis, a significant difference was demonstrated between the candidate and reference methods for the high spiking level with raw ground pork and brie cheese. For all other matrixes and the low spiked levels for raw ground pork and brie cheese, no significant difference by POD was seen between the two methods during the study.

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