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

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.

2013 ◽  
Vol 96 (3) ◽  
pp. 532-541
Author(s):  
Olga Petrauskene ◽  
Yan Cao ◽  
Patrick Zoder ◽  
Lily Y Wong ◽  
Priya Balachandran ◽  
...  

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.


2013 ◽  
Vol 96 (2) ◽  
pp. 225-228 ◽  
Author(s):  
DeAnn L Benesh ◽  
Erin S Crowley ◽  
Patrick M Bird

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.


2013 ◽  
Vol 96 (2) ◽  
pp. 218-224
Author(s):  
DeAnn L Benesh ◽  
Erin S Crowley ◽  
Patrick M Bird

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.


2013 ◽  
Vol 96 (3) ◽  
pp. 516-523
Author(s):  
Wendy F Lauer ◽  
Jean-Philippe Tourniaire

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.


2013 ◽  
Vol 96 (3) ◽  
pp. 508-515
Author(s):  
Wendy F Lauer ◽  
Jean-Philippe Tourniaire

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.


2011 ◽  
Vol 94 (4) ◽  
pp. 1106-1116 ◽  
Author(s):  
Priya Balachandran ◽  
Yanxiang Cao ◽  
Lily Wong ◽  
Manohar R Furtado ◽  
Olga V Petrauskene ◽  
...  

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.


1997 ◽  
Vol 80 (4) ◽  
pp. 806-824 ◽  
Author(s):  
Michael T Knight ◽  
Melissa C Newman ◽  
M Joseph Benzinger ◽  
Karen L Neufang ◽  
James R Agin ◽  
...  

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.


2011 ◽  
Vol 94 (5) ◽  
pp. 1467-1480
Author(s):  
Rebecca Hoerner ◽  
Jill Feldpausch ◽  
R Lucas Gray ◽  
Stephanie Curry ◽  
Zahidul Islam ◽  
...  

Abstract Reveal Salmonella 2.0 is an improved version of the original Reveal Salmonella lateral flow immunoassay and is applicable to the detection of Salmonella enterica serogroups A–E in a variety of food and environmental samples. A Performance Tested MethodSM validation study was conducted to compare performance of the Reveal 2.0 method with that of the U.S. Department of Agriculture-Food Safety and Inspection Service or U.S. Food and Drug Administration/Bacteriological Analytical Manual reference culture methods for detection of Salmonella spp. in chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, raw shrimp, a ready-to-eat meal product, dry pet food, ice cream, spinach, cantaloupe, peanut butter, stainless steel surface, and sprout irrigation water. In a total of 17 trials performed internally and four trials performed in an independent laboratory, there were no statistically significant differences in performance of the Reveal 2.0 and reference culture procedures as determined by Chi-square analysis, with the exception of one trial with stainless steel surface and one trial with sprout irrigation water where there were significantly more positive results by the Reveal 2.0 method. Considering all data generated in testing food samples using enrichment procedures specifically designed for the Reveal method, overall sensitivity of the Reveal method relative to the reference culture methods was 99%. In testing environmental samples, sensitivity of the Reveal method relative to the reference culture method was 164%. For select foods, use of the Reveal test in conjunction with reference method enrichment resulted in overall sensitivity of 92%. There were no unconfirmed positive results on uninoculated control samples in any trials for specificity of 100%. In inclusivity testing, 102 different Salmonella serovars belonging to serogroups A–E were tested and 99 were consistently positive in the Reveal test. In exclusivity testing of 33 strains of non-salmonellae representing 14 genera, 32 were negative when tested with Reveal following nonselective enrichment, and the remaining strain was found to be substantially inhibited by the enrichment media used with the Reveal method. Results of ruggedness testing showed that the Reveal test produces accurate results even with substantial deviation in sample volume or device development time.


2009 ◽  
Vol 92 (6) ◽  
pp. 1885-1889 ◽  
Author(s):  
Charlotte Lindhardt ◽  
Holger Schönenbrücher ◽  
Jörg Slaghuis ◽  
Andreas Bubert ◽  
Rolf Ossmer ◽  
...  

Abstract Singlepath Salmonella is an immunochromatographic (lateral flow) assay for the presumptive qualitative detection of Salmonella spp. in food. A previous AOAC Performance Tested MethodSM study evaluated Singlepath Salmonella as an effective method for the detection of Salmonella spp. in the following selected foods: dried skimmed milk, black pepper, dried pet food, desiccated coconut, cooked peeled frozen prawns, raw ground beef, and raw ground turkey. In this Emergency Response Validation extension, creamy peanut butter was inoculated with S. enterica. ser. Typhimurium. For low contamination level (1.08 CFU/25 g), a Chi-square value of 0.5 indicated that there was no significant difference between Singlepath Salmonella and the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) reference method. For high-level and uninoculated control there was 100 agreement between the methods.


2003 ◽  
Vol 86 (2) ◽  
pp. 340-354 ◽  
Author(s):  
Denise Hughes ◽  
Angela Dailianis ◽  
Louise Duncan ◽  
Julie Briggs ◽  
Deborah A McKintyre ◽  
...  

Abstract A collaborative study was conducted to validate new enrichment methods for the TECRA Listeria Visual Immunoassay (TLVIA). These new methods incorporate a newly formulated medium, TECRA Listeria Enrichment Broth, which does not contain the highly toxic antifungal agent, cycloheximide. The new procedures will provide an alternative to the enrichment procedures described in AOAC Method 995.22. Three food types (raw ground beef, lettuce, and ice cream) were analyzed in the United States, and 2 food types (cooked turkey and cooked fish fillets) were analyzed in Australasia. Thirty collaborators participated in the study, 16 in Australasia and 14 in the United States. With the exception of one batch of ground beef, comparison of the proportion of positive test portions (p ≥ 0.05) showed no significant difference between the TLVIA and the reference method for the 5 foods at 3 inoculation levels. For the one batch of naturally contaminated raw ground beef, the TLVIA gave significantly more confirmed positive results than the reference method.


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