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.

Method Modification Study for the Thermo Scientific SureTect™ Listeria Species Assay–Matrix Extension

2016 ◽  
Vol 99 (2) ◽  
pp. 417-427
Author(s):  
Jonathan Cloke ◽  
Katharine Evans ◽  
David Crabtree ◽  
Annette Hughes ◽  
Helen Simpson ◽  
...  
Keyword(s):  
Reference Method ◽  
Probability Of Detection ◽  
Pcr Assay ◽  
Listeria Species ◽  
Significant Difference ◽  
Thermo Fisher Scientific ◽  
Pork Sausages ◽  
The University ◽  
Matrix Extension ◽  
Fisher Scientific

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. The assay was originally certified as Performance Tested MethodsSM (PTM) 071304 in 2013. This report details the method modification study undertaken to extend the performance claims of the assay for matrixes of raw ground turkey, raw ground pork, bagged lettuce, raw pork sausages, pasteurized 2% fat milk, raw cod, pasteurized brie cheese, and ice cream. The method modification study was conducted using the AOAC Research Institute (RI) PTM program to validate the SureTect PCR assay in comparison to the reference method detailed in ISO 11290-1:1996 including amendment 1:2004. All matrixes were tested by Thermo Fisher Scientific (Basingstoke, United Kingdom). In addition, three matrixes (raw cod, bagged lettuce, and pasteurized brie cheese) 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, there was no significant difference in the performance between the SureTect assay and the International Organization for Standardization reference method for any of the matrixes analyzed in this study.

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.

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.

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.

Comparative Evaluation of Veriflow®Listeria monocytogenes to USDA and AOAC Culture Based Methods for the Detection of Listeria monocytogenes in Food

2015 ◽  
Vol 98 (5) ◽  
pp. 1325-1334
Author(s):  
Adam C Joelsson ◽  
Ashley S Brown ◽  
Amrita Puri ◽  
Martin P Keough ◽  
Zara E Gaudioso ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Pcr Amplification ◽  
Reference Method ◽  
Probability Of Detection ◽  
Complex Data ◽  
Detection Analysis ◽  
Environmental Surfaces ◽  
Study Results ◽  
Significant Difference ◽  
Inspection Service

Abstract Veriflow®Listeria monocytogenes (LM) is a molecular based assay for the presumptive detection of Listeria monocytogenes from environmental surfaces, dairy, and ready-to-eat (RTE) food matrixes (hot dogs and deli meat). The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post PCR amplification and requires only 24 h of enrichment for maximum sensitivity. The Veriflow LM 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 LM method to detect low levels of artificially inoculated L. monocytogenes in seven distinct environmental and food matrixes. In each unpaired reference comparison study, probability of detection analysis indicated no significant difference between the Veriflow LM method and the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook 8.08 or AOAC 993.12 reference method. Fifty strains of L. monocytogenes were detected in the inclusivity study, while 39 nonspecific organisms were undetected in the exclusivity study. The study results show that Veriflow LM is a sensitive, selective, and robust assay for the presumptive detection of L. monocytogenes sampled from environmental, dairy, or RTE (hot dogs and deli meat) food matrixes.

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.

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 GENE-UP ® Listeria monocytogenes Method for the Detection of Listeria monocytogenes in a Variety of Foods and Select Environmental Surfaces: Collaborative Study, First Action 2019.11

2020 ◽  
Author(s):  
Ronald Johnson ◽  
John Mills ◽  
Jean-Louis Pittet ◽  
Olivier Mathia ◽  
Patrick Bird ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Collaborative Study ◽  
Control Level ◽  
Reference Method ◽  
Probability Of Detection ◽  
Cottage Cheese ◽  
The European Union ◽  
Inoculum Level ◽  
Test Portion ◽  
Environmental Surfaces

Abstract Background The GENE-UP®Listeria monocytogenes 2 (LMO 2) assay (Performance Tested MethodSM 121804) uses real-time PCR technology and a proprietary detection platform, the GENE-UP® Thermocycler, to detect Listeria monocytogenes in a variety of foods and environmental surfaces. Objective The purpose of this validation was to evaluate the method’s interlaboratory performance and submit the result to AOAC INTERNATIONAL for adoption as First Action Official MethodSM for the detection of Listeria monocytogenes in a variety of foods and select environmental surfaces. Method The GENE-UP® method was evaluated in a multi-laboratory study as part of the AFNOR NF VALIDATION certification process using unpaired test portions for one food matrix, full-cream goat milk cottage cheese (8.4% fat). The candidate method was compared to the ISO 11290-1/Amd.1:2004 reference method. Sixteen participants from 15 laboratories throughout the European Union participated. Three levels of contamination were evaluated: a non-inoculated control level (0 CFU/test portion), a low inoculum level (∼2 CFU/test portion) and a high inoculum level (∼10 CFU/test portion). Data from the study were analyzed according to the Probability of Detection (POD) statistical model as presented in the AOAC validation guidelines. Results The dLPODC values with 95% confidence interval for each comparison were; -0.02 (-0.07, 0.03), -0.08 (-0.31, 0.16) and 0.00 (-0.03, 0.03) for the non-inoculated, low and high contamination levels respectively. Conclusion The dLPODC results demonstrate no difference in performance between the candidate method and reference method for the matrix evaluated. Highlights The GENE-UP LMO method demonstrated accuracy and precision in detecting and discerning L. monocytogenes from other Listeria species.

scholarly journals Comparative Evaluation of the Thermo Fisher TaqPath TM COVID-19 Combo Kit with the Cepheid Xpert ® Xpress SARS-CoV-2 Assay for Detecting SARS-CoV-2 in Nasopharyngeal Specimens

2021 ◽  
Author(s):  
Paul A. Granato ◽  
Simon R. Kimball ◽  
Brenda R. Alkins ◽  
Deirdre C. Cross ◽  
Melissa M. Unz
Keyword(s):  
Molecular Diagnostics ◽  
Comparative Evaluation ◽  
Sanger Sequencing ◽  
Experimental Validation ◽  
High Complexity ◽  
Pcr Assay ◽  
Clinical Specimens ◽  
Percent Agreement ◽  
Thermo Fisher Scientific ◽  
Fisher Scientific

Abstract Purpose With over 50 SARS-CoV-2 gene amplification assays that have been EUA cleared with minimal experimental validation performed, it is likely that not all of these assays are comparable in their ability to detect SARS-CoV-2 in clinical specimens. Thermo Fisher Scientific is a relatively new company in the molecular diagnostics field and the purpose of this study was to compare the performance of the Thermo Fisher TaqPath™ Combo Kit with an established test, the Cepheid Xpert® Xpress SARS-CoV-2 assay, for its ability to detect SARS-CoV-2 in nasopharyngeal specimens.Methods A total of 300 randomly selected nasopharyngeal specimens were evaluated and tested by the TaqPath and GeneXpert assays. Discordant test specimens were arbitrated by performing an alternative PCR assay and Sanger sequencing.Results The TaqPath assay had a 96.7% and 99.6% positive and negative agreement respectively when compared to the Xpert Xpress test. However, after test arbitration, the three discordant specimens were arbitrated in favor of the TaqPath assay producing a positive and negative percent agreement of 100% for the TaqPath Combo Kit while the Xpress SARS-CoV-2 assay had a positive and negative percent agreement of 98.3% and 99.2% respectively.Conclusions The TaqPath Combo Kit is a high complexity assay that compares favorably with the Xpert Xpress test and can be reliably used for the detection of SARS-CoV-2 in nasopharyngeal specimens.

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