InstantLabs®Salmonella Species Detection Method: Matrix Extension

2014 ◽  
Vol 97 (6) ◽  
pp. 1585-1591
Author(s):  
Neil Sharma ◽  
Lauren Bambusch ◽  
Thu Le ◽  
Amit Morey ◽  
Melinda Hayman ◽  
...  
Keyword(s):  
Ground Beef ◽  
Reference Method ◽  
Test Method ◽  
Chicken Breast ◽  
Escherichia Coli O157 ◽  
Test Portion ◽  
Screen Result ◽  
Low Levels ◽  
Matrix Extension ◽  
Better Than

Abstract The performance of InstantLabs®Salmonella Species Food Safety Kit to detect Salmonella in four food matrixes was validated against the International Organization for Standardization (ISO) reference method 6579:2002. The matrixes (raw ground beef, raw chicken breast, raw ground chicken, and lettuce) were inoculated with low levels of Salmonella (<1 CFU/test portion) to generate fractional positives (5–15) in 20 inoculated samples. These matrixes were co-inoculated with Escherichia coli O157:H7 at two to five times the level of Salmonella. Samples were validated using 375 g (meat) or 25 g (lettuce and poultry) test portions enriched in FASTGRO™ SE at 42 ± 1°C for 12 h and 10 h, respectively. All samples were confirmed using the ISO reference method, regardless of initial screen result. The InstantLabs test method was shown to perform as well as or better than the reference method for the detection of Salmonella species in ground beef, chicken breast, ground chicken, and lettuce. Inclusivity and exclusivity testing revealed no false negatives among the 100 Salmonella serovars and no false positives among the 30 non-Salmonella species examined, respectively.

InstantLabs®Salmonella Species Food Safety Kit

2014 ◽  
Vol 97 (6) ◽  
pp. 1576-1584
Author(s):  
Neil Sharma ◽  
Lauren Bambusch ◽  
Thu Le ◽  
Melinda Hayman ◽  
Sergio J Montez
Keyword(s):  
Food Safety ◽  
Wheat Flour ◽  
Enrichment Culture ◽  
Hold Time ◽  
Reference Method ◽  
Test Method ◽  
Test Portion ◽  
Screen Result ◽  
Oat Flour ◽  
Better Than

Abstract The InstantLabs®Salmonella Species Food Safety Kit was validated against the International Organization for Standardization (ISO) reference method 6579:2002 for the detection of Salmonella species. The matrixes (unprocessed rolled oats, wheat flour, and oat flour) were inoculated with 1 CFU/test portion of Salmonella to generate fractional positives (5–15) in 20 inoculated samples. The matrixes were co-inoculated with Escherichia coli O157:H7 at 2–5 times the level of Salmonella to demonstrate the potential for using the same enrichment culture in the future to detect of multiple organisms. Samples were validated using 750 g test portion enriched in FASTGRO SE at 42 ± 1°C for 16–20 h. All samples were confirmed using the ISO reference method, regardless of initial screen result. The InstantLabs test method performed as well as or better than the reference method for the detection of Salmonella species in unprocessed rolled oats, wheat flour, and oat flour. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 100 Salmonella serovars and 30 non-Salmonella species examined. Finally, the method was shown to be robust when variations to enrichment time, DNA extract hold time, and DNA volume were varied (data not shown).

InstantLabs®E. coli O157 Food Safety Kit

2015 ◽  
Vol 98 (1) ◽  
pp. 71-81
Author(s):  
Neil Sharma ◽  
Lauren Bambusch ◽  
Apala Upadhyay ◽  
Thu Le ◽  
Chris Lopez ◽  
...  
Keyword(s):  
Food Safety ◽  
Enrichment Culture ◽  
Reference Method ◽  
Test Method ◽  
Romaine Lettuce ◽  
Test Portion ◽  
E Coli ◽  
Screen Result ◽  
Block Time ◽  
Better Than

Abstract The InstantLabs®E. coli O157 Food Safety Kit was validated against the International Organization for Standardizationreference method 16654 for the detection of Escherichia coli O157. The matrixes, raw ground beef, raw beef trim, Romaine lettuce, pasteurized apple juice, and raw ground chicken, were inoculated with appropriate CFU/test portion of E. coli O157 to generate fractional positives (5–15) in 20 inoculated samples. The matrixeswere co-inoculated with Salmonella at 2–5 times the level of E. coli O157 to demonstrate the potential for using the same enrichment culture for the detection of multiple organisms. Samples were enriched in prewarmed FASTGRO SE broth at 42 ± 1°C for 10–20 h. All samples were confirmed using the ISO reference method, regardless of initial screen result. The InstantLabs test method performed as well as or better than the reference method for the detection of E. coli O157 in all tested samples. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 50 E. coli O157 serovars and 30 non-E. coli O157 species examined. Finally, the method was shown to be robust when variations were applied to enrichment time, volume for DNA extraction, and heat block time.

scholarly journals Modification and Matrix Extension of the Bio-Rad iQ-Check E. coli O157:H7, STEC VirX, and STEC SerO Test Kits for the Detection of Shiga Toxin–Producing Escherichia coli (STEC) and Escherichia coli O157 From a Single Enrichment

2020 ◽  
Vol 103 (1) ◽  
pp. 161-175
Author(s):  
Dane Brooks ◽  
Benjamin Bastin ◽  
Erin Crowley ◽  
James Agin ◽  
Mike Clark ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Ground Beef ◽  
Reference Method ◽  
Chicken Breast ◽  
E Coli ◽  
Free Dna ◽  
Target Analytes ◽  
Matrix Extension

Abstract Background: The iQ-Check Real-Time PCR kits use PCR technology based on gene amplification and detection by a real-time PCR thermalcycler for the detection of target analytes in select food matrices. The iQ-Check E. coli O157:H7 [Performance Tested MethodSM (PTM) 020801] and STEC VirX and STEC SerO (combined PTM 121203) methods were previously validated for different matrices under different enrichment schemes. Objective: To modify the current iQ-Check E. coli O157:H7 Kit for the detection of Escherichia coli O157:H7 from 25 to 375 g for raw ground beef (17% fat), raw beef trim, and fresh spinach. In addition, a matrix extension was validated for iQ-Check E. coli O157:H7 for raw chicken breast without skin (25 g), raw chicken thigh with skin (25 g), mechanically separated chicken (25 g), and raw ground pork (25 g). The study also included the modification of the iQ-Check STEC VirX and SerO Kits for the detection of non-O157 Shiga toxin–producing E. coli (STEC) for raw ground beef (375 g), raw beef trim (375 g), and fresh spinach (375 g) from STEC Enrichment Broth to buffered peptone water (BPW). All tests were carried out at 8–22 h (10–22 h for fresh spinach). Methods: Ground beef, beef trim, and spinach were co-inoculated with E. coli O157:H7, non-O157 STECs, and Salmonella spp. and analyzed for E. coli O157:H7 and non-O157 STECs after an 8-22 h enrichment in BPW for the beef matrices and after a 10–22 h enrichment in BPW for spinach. The chicken matrices were inoculated with E. coli O157:H7 only and analyzed after an 8–22 h enrichment in BPW. The iQ-Check Free DNA Removal Solution workflow was utilized for all matrices. Confirmations at the 22 h time point and method comparisons were conducted with the appropriate reference method as outlined in the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 4A or the U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook Chapters 5.09 and 5B.05. For the iQ-Check STEC VirX and STEC SerO Kits, inclusivity and exclusivity were also performed. Results: The two inclusivity and exclusivity evaluations indicated that the test methods can accurately detect the target analytes and correctly excluded nontarget organisms after 8 h of enrichment. In the method comparison study, the iQ-Check E. coli O157:H7 and STEC VirX and STEC SerO test kits demonstrated no statistically significant differences between candidate and reference method results or between presumptive and confirmed results for all food matrices analyzed and the two time points (8 or 10 and 22 h). Both time points produced the same results, with no discrepancies. Conclusions: The iQ-Check real-time PCR kits are effective methods for the detection of E. coli O157 and non-O157 STECs (both the virulence factors and the O groups) from raw ground beef, raw beef trim, and fresh spinach in 375 g samples enriched in BPW for 8–22 h (10–22 h for fresh spinach). In addition, the iQ-Check E. coli O157 Kit is effective in detecting E. coli O157 in 25 g samples of raw chicken breast without skin, raw chicken thigh with skin, mechanically separated chicken, and raw ground pork. The iQ-Check test kits allow the end user to pair enrichments for multiple target analytes, allowing the user to prepare a single enrichment and perform a single DNA extraction. The Free DNA Removal Solution removes free DNA from samples prior to PCR analysis, protecting DNA from intact and living cells. Highlights: The method modifications were granted based on the data collected.

Distribution Patterns of Escherichia coli O157:H7 in Ground Beef Produced by a Laboratory-Scale Grinder†

2002 ◽  
Vol 65 (12) ◽  
pp. 1894-1902 ◽  
Author(s):  
ROLANDO A. FLORES ◽  
MARK L. TAMPLIN
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Distribution Patterns ◽  
Small Scale ◽  
Escherichia Coli O157 ◽  
Inoculum Level ◽  
E Coli ◽  
Specific Distribution ◽  
Low Levels ◽  
G Prior

This study determined the distribution patterns of Escherichia coli O157:H7 in ground beef when a contaminated beef trim was introduced into a batch of uncontaminated beef trims prior to grinding in a small-scale laboratory grinder. A beef trim (15.3 ± 2 g) was inoculated with a rifampicin-resistant strain of E. coli O157:H7 (E. coli O157:H7rif) and introduced into a stream of noncontaminated beef (322 ± 33 g) prior to grinding. Seven inoculum levels (6, 5, and 4 total log CFU [high]; and 3, 2, 1, and 0 total log CFU [low]) were studied in triplicate. E. coli O157:H7rif was not detected in 3.1 to 43% of the ground beef inoculated with the high levels or in 3.4 to 96.9% of the ground beef inoculated with the low levels. For all inoculum levels studied, the five ground beef fractions (each 7.8 ± 0.6 g) with the highest pathogen levels accounted for 59 to 100% of the total pathogens detected. For all inoculum levels, there was a linear relationship between the quantity of ground beef containing E. coli O157:H7rif and the inoculum level. The quantity of E. coli O157:H7rif in the beef remaining in the grinder was proportional to the inoculum level and was related to the location in the grinder. Different components of the grinder accumulated E. coli O157:H7rif in different quantities, with the most significant accumulation being in the nut (collar) that attaches the die to the blade. This study determined specific distribution patterns of E. coli O157:H7rif after the grinding of a contaminated beef trim along with uncontaminated trims, and the results indicate that the grinding operation should be regarded as a means of distribution of microbial contamination in risk analyses of ground beef operations.

scholarly journals VIDAS Salmonella (SLM) Assay Method EasySLM with ChromID Salmonella (SM2) Agar

2009 ◽  
Vol 92 (6) ◽  
pp. 1861-1864 ◽  
Author(s):  
Ronald Johnson ◽  
John Mills ◽  
Judith Coln-Reveles ◽  
Thomas Hammack
Keyword(s):  
Peanut Butter ◽  
Reference Method ◽  
Black Pepper ◽  
Assay Method ◽  
Most Probable Number ◽  
Chicken Breast ◽  
Probable Number ◽  
Pork Sausage ◽  
Selective Enrichment ◽  
Matrix Extension

Abstract A method modification study was conducted for the VIDAS Salmonella (SLM) assay (AOAC Performance Tested MethodSM 020901) using the EasySLM method to validate a matrix extension for peanut butter. The VIDAS EasySLM method is a simple enrichment procedure compared to traditional Salmonella methods, requiring only pre-enrichment and a single selective enrichment media, Salmonella Xpress 2 (SX2) broth. SX2 replaces the two selective broths in traditional methods and eliminates the M broth transfer, incubation, and subsequent pooling of M broths prior to VIDAS assay. The validation study was conducted under the AOAC Research Institute Emergency Response Validation program. VIDAS SLM was compared to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) method for detection of S. enterica ser. Typhimurium in peanut butter. All peanut butter samples were prepared, blind-coded, and shipped to the method developers' laboratory by Q Laboratories. In addition, Q Laboratories performed most probable number and reference method analyses on peanut butter samples. The VIDAS EasySLM ChromID Salmonella (SM2) Agar was previously validated in the Performance Tested Methods program for the detection of Salmonella in roast beef, raw ground pork, turkey, pork sausage, raw chicken breast, dry pet food, whole milk, ice cream, bagged spinach, shrimp (raw, peeled), raw cod, spent irrigation water, pecans, peanut butter, dry pasta, cake mix, ground black pepper, nonfat dry milk, liquid eggs, cantaloupe, and orange juice. In the matrix extension study for peanut butter, the VIDAS EasySLM method was shown to be equivalent to the appropriate reference culture procedure using both buffered peptone water pre-enrichment and the FDA-BAM lactose pre-enrichment in the two-step enrichment method with SX2 media. The current study extends the validation to include peanut butter.

Comparative Evaluation of a Phage Protein Ligand Assay with Real-Time PCR and a Reference Method for the Detection of Escherichia coli O157:H7 in Raw Ground Beef and Trimmings

2011 ◽  
Vol 74 (1) ◽  
pp. 6-12 ◽  
Author(s):  
F. SAVOYE ◽  
P. FENG ◽  
C. ROZAND ◽  
M. BOUVIER ◽  
A. GLEIZAL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Ground Beef ◽  
Reference Method ◽  
Enterohemorrhagic Escherichia Coli ◽  
Detection Methods ◽  
Escherichia Coli O157 ◽  
Rt Pcr ◽  
Raw Meat ◽  
E Coli

Enterohemorrhagic Escherichia coli O157:H7 is an important pathogen associated with infections caused by consumption of undercooked raw meat. Sensitive and rapid detection methods for E. coli O157:H7 are essential for the meat industry to ensure a safe meat supply. This study was conducted to compare the sensitivity of the VIDAS ultraperformance E. coli test (ECPT UP) with a noncommercial real-time (RT) PCR method and the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) reference method for detecting E. coli O157:H7 in raw ground beef. Optimal enrichment times and the efficacy of testing different types of raw meat, either as individual samples (25 g) or as composites (375 g), were examined. For 25-g samples of each type of raw ground beef tested, 6 h of enrichment was sufficient for both the VIDAS ECPT UP and RT-PCR methods, but for 375-g samples, 24 h of enrichment was required. Both the VIDAS ECPT UP and RT-PCR methods produced results similar to those obtained with the USDA-FSIS reference method after 18 to 24 h of enrichment. The primer specificity of the RT-PCR assay and the highly specific phage ligand used in the VIDAS ECPT UP for target recognition enabled the detection of low levels of E. coli O157:H7 in 25 g of various types of raw ground beef. The tests also allowed the detection of E. coli O157:H7 in composite raw ground beef and trimmings in samples of up to 375 g.

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.

Validation of the Thermo Scientific SureTect Escherichia coli O157:H7 Real-Time PCR Assay for Raw Beef and Produce Matrixes

2015 ◽  
Vol 98 (5) ◽  
pp. 1301-1314 ◽  
Author(s):  
Jonathan Cloke ◽  
Erin Crowley ◽  
Patrick Bird ◽  
Ben Bastin ◽  
Jonathan Flannery ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Real Time ◽  
Real Time Pcr ◽  
Apple Juice ◽  
Ground Beef ◽  
Reference Method ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
E Coli

Abstract The Thermo Scientific™ SureTect™ Escherichia coli O157:H7 Assay is a new real-time PCR assay which has been validated through the AOAC Research Institute (RI) Performance Tested MethodsSM program for raw beef and produce matrixes. This validation study specifically validated the assay with 375 g 1:4 and 1:5 ratios of raw ground beef and raw beef trim in comparison to the U.S. Department of Agriculture, Food Safety Inspection Service, Microbiology Laboratory Guidebook (USDS-FSIS/MLG) reference method and 25 g bagged spinach and fresh apple juice at a ratio of 1:10, in comparison to the reference method detailed in the International Organization for Standardization 16654:2001 reference method. For raw beef matrixes, the validation of both 1:4 and 1:5 allows user flexibility with the enrichment protocol, although which of these two ratios chosen by the laboratory should be based on specific test requirements. All matrixes were analyzed by Thermo Fisher Scientific, Microbiology Division, Vantaa, Finland, and Q Laboratories Inc, Cincinnati, Ohio, in the method developer study. Two of the matrixes (raw ground beef at both 1:4 and 1:5 ratios) and bagged spinach were additionally analyzed in the AOAC-RI controlled independent laboratory study, which was conducted by Marshfield Food Safety, Marshfield, Wisconsin. Using probability of detection statistical analysis, no significant difference was demonstrated by the SureTect kit in comparison to the USDA FSIS reference method for raw beef matrixes, or with the ISO reference method for matrixes of bagged spinach and apple juice. Inclusivity and exclusivity testing was conducted with 58 E. coli O157:H7 and 54 non-E. coli O157:H7 isolates, respectively, which demonstrated that the SureTect assay was able to detect all isolates of E. coli O157:H7 analyzed. In addition, all but one of the nontarget isolates were correctly interpreted as negative by the SureTect Software. The single isolate giving a positive result was an E. coli O157:NM isolate. Nonmotile isolates of E. coli O157 have been demonstrated to still contain the H7 gene; therefore, this result is not unexpected. Robustness testing was conducted to evaluate the performance of the SureTect assay with specific deviations to the assay protocol, which were outside the recommended parameters and which are open to variation. This study demonstrated that the SureTect assay gave reliable performance. A final study to verify the shelf life of the product, under accelerated conditions was also conducted.

Evaluation of the 3M™ Petrifilm™ Salmonella Express System for the Detection of Salmonella Species in Selected Foods: Collaborative Study

2014 ◽  
Vol 97 (6) ◽  
pp. 1563-1575 ◽  
Author(s):  
Patrick Bird ◽  
Jonathan Flannery ◽  
Erin Crowley ◽  
James Agin ◽  
David Goins ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Control Level ◽  
Ground Beef ◽  
Reference Method ◽  
Probability Of Detection ◽  
Isolation And Identification ◽  
Inoculum Level ◽  
Test Portion ◽  
Dog Food ◽  
The U.S

Abstract The 3M™ Petrifilm™Salmonella Express (SALX) System is a simple, ready-to-use chromogenic culture medium system for the rapid qualitative detection and biochemical confirmation of Salmonella spp. in food and food process environmental samples. The 3M Petrifilm SALX System was compared using an unpaired study design in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.07 (2013) Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products and Carcass and Environmental Sponges for raw ground beef and the U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA/BAM) Chapter 5, Salmonella (2011) reference method for dry dog food following the current AOAC validation guidelines. For this study, a total of 17 laboratories located throughout the continental United States evaluated 1872 test portions. For the 3M Petrifilm SALX System, raw ground beef was analyzed using 25 g test portions, and dry dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. The two matrices were 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). Each inoculation level was statistically analyzed using the probability of detection statistical model. For the raw ground beef and dry dog food test portions, no significant differences at the 95% confidence interval were observed in the number of positive samples detected by the 3M Petrifilm SALX System versus either the USDA/FSIS-MLG or FDA/BAM methods.

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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