SAS™ Molecular Tests Salmonella Detection Kit

2014 ◽  
Vol 97 (3) ◽  
pp. 808-819 ◽  
Author(s):  
Chandra Bapanpally ◽  
Laura Montier ◽  
Shah Khan ◽  
Akif Kasra ◽  
Sharon L Brunelle
Keyword(s):  
Hold Time ◽  
Ground Beef ◽  
Department Of Agriculture ◽  
Test Portion ◽  
Method Performance ◽  
Molecular Tests ◽  
Amplification Method ◽  
Chicken Carcass ◽  
Inspection Service ◽  
The U.S

Abstract The SAS™ Molecular tests Salmonella Detection method, a Loop-mediated Isothermal Amplification method, performed as well as or better than the U.S. Department of Agriculture-Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, ground turkey, chicken carcass rinses, bagged mixed lettuce, and fresh spinach. The ground beef (30% fat, 25 g test portion), poultry matrixes and leafy greens were validated in a 6–7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16–20 h enrichment. The method performance for meat and leafy green matrixes was shown to be acceptable under conditions of co-enrichment with Escherichia coli O157. Thus, after a short 6–7 h co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 100 Salmonella serovars and 30 non- Salmonella species examined. The method was shown to be robust when enrichment time, DNA extract hold time, and DNA volume were varied.

SAS™ Molecular tests Escherichia coli O157 Detection Kit

2014 ◽  
Vol 97 (3) ◽  
pp. 798-807 ◽  
Author(s):  
Chandra Bapanpally ◽  
Laura Montier ◽  
Shah Khan ◽  
Akif Kasra ◽  
Sharon L Brunelle
Keyword(s):  
Escherichia Coli ◽  
Hold Time ◽  
Method Comparison ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Test Portion ◽  
Method Performance ◽  
E Coli ◽  
Molecular Tests ◽  
The U.S

Abstract The SAS™ Molecular tests Escherichia coli O157 Detection method, a loop-mediated isothermal amplification method, performed as well as or better than the U.S. Department of Agriculture, Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, bagged mixed lettuce, and fresh spinach. Ground beef (30% fat, 25 g test portion) was validated for 7–8 h enrichment, leafy greens were validated in a 6–7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16–20 h enrichment. The method performance for meat and leafy green matrixes was also shown to be acceptable under conditions of co-enrichment with Salmonella. Thus, after a short co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. The SAS Molecular tests Salmonella Detection Kit was validated using the same test portions as for the SAS Molecular tests E. coli O157 Detection Kit and those results are presented in a separate report. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 50 E. coli O157 strains, including H7 and non-motile strains, and 30 non-E. coli O157 strains examined. Finally, the method was shown to be robust when variations to DNA extract hold time and DNA volume were varied. The method comparison and robustness data suggest a full 7 h enrichment time should be used for 25 g ground beef test portions.

Validation of the Reveal® 2.0 Group D1 Salmonella Test for Detection of Salmonella Enteritidis in Raw Shell Eggs and Poultry-Associated Matrixes

2016 ◽  
Vol 99 (4) ◽  
pp. 1017-1024 ◽  
Author(s):  
Mark Mozola ◽  
Preetha Biswas ◽  
Ryan Viator ◽  
Emily Feldpausch ◽  
Debra Foti ◽  
...  
Keyword(s):  
Salmonella Enteritidis ◽  
Probability Of Detection ◽  
Poultry Feed ◽  
Operating Parameters ◽  
Department Of Agriculture ◽  
Shell Eggs ◽  
Chicken Carcass ◽  
Inspection Service ◽  
The U.S ◽  
Culture Procedure

Abstract A study was conducted to assess the performance of the Reveal® 2.0 Group D1 Salmonella lateral flow immunoassay for use in detection of Salmonella Enteritidis (SE) in raw shell eggs and poultry-associated matrixes, including chicken carcass rinse and poultry feed. In inclusivity testing, the Reveal 2.0 test detected all 37 strains of SE tested. The test also detected all but one of 18 non-Enteritidis somatic group D1 Salmonella serovars examined. In exclusivity testing, none of 42 strains tested was detected. The exclusivity panel included Salmonella strains of somatic groups other than D1, as well as strains of other genera of Gram-negative bacteria. In matrix testing, performance of the Reveal 2.0 test was compared to that of the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference culture procedure for chicken carcass rinse and to that of the U.S. Food and Drug Administration Bacteriological Analytical Manual for raw shell eggs and poultry feed. For all matrixes evaluated, there were no significant differences in the ability to detect SE when comparing the Reveal 2.0 method and the appropriate reference culture procedure as determined by probability of detection statistical analysis. The ability of the Reveal 2.0 test to withstand modest perturbations to normal operating parameters was examined in robustness experiments. Results showed that the test can withstand deviations in up to three operating parameters simultaneously without significantly affecting performance. Real-time stability testing of multiple lots of Reveal 2.0 devices established the shelf life of the test device at 16 months postmanufacture.

Crystal Diagnostics MultiPath System™

2014 ◽  
Vol 97 (6) ◽  
pp. 1592-1600 ◽  
Author(s):  
Curtis H Stumpf ◽  
Weidong Zhao ◽  
Brian Bullard ◽  
Christine Ammons ◽  
Karl I Devlin ◽  
...  
Keyword(s):  
Liquid Crystal ◽  
Laboratory Tests ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Department Of Agriculture ◽  
E Coli ◽  
Independent Laboratory ◽  
Diagnostics System ◽  
Inspection Service ◽  
The U.S

Abstract The Crystal Diagnostics MultiPath System™ provides rapid detection of Escherichia coli O157 in fresh raw ground beef, raw beef trim, and spinach. The Crystal Diagnostics system combines patented Liquid Crystal technology with antibody-coated paramagnetic microspheres to selectively capture and detect E. coli O157 in food matrixes. This is the only liquid crystal-based biosensor commercially available for the detection of pathogens. The Crystal Diagnostics system expeditiously provides the sensitivity and accuracy of the U.S. Department of Agriculture Food Safety Inspection Service (USDA-FSIS) and the U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA-BAM) methods for detecting as low as one CFU of E. coli O157 per 375 g of raw ground beef and raw beef trim, or 200 g of raw spinach. An internal inclusivity validation demonstrated detection of all 50 tested strains of E. coli O157. The internal and independent laboratory tests demonstrate that the method is rapid and sensitive for detecting of E. coli O157 in fresh raw ground beef, beef trim, and spinach.

Comparative Validation Study to Demonstrate the Equivalence of an Alternate Next-Day Enrichment Protocol for VIP® Gold for Salmonella Method to Culture Methods for the Detection of Salmonella in Selected Foods and Environmental Surfaces

2019 ◽  
Vol 102 (3) ◽  
pp. 815-827
Author(s):  
David E Kerr ◽  
George Shen ◽  
Andrew H Lienau ◽  
Mandeep Kaur ◽  
Amy L Immermann ◽  
...  
Keyword(s):  
Enterohemorrhagic Escherichia Coli ◽  
Official Method ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Environmental Surfaces ◽  
Chicken Carcass ◽  
Significant Difference ◽  
Enrichment Protocol ◽  
Inspection Service ◽  
The U.S

Abstract Background: VIP® Gold for Salmonella is a lateral flow immunodetection device that was validated by AOAC in 1999 as Official Method of Analysis 999.09. It was improved upon in 2009 by introducing gold colloid as the detection method. Objective: A simple next-day enrichment protocol using modified enterohemorrhagic Escherichia coli media was developed for the VIP Gold for Salmonella to improve the time-to-results and laboratory work flow. Methods: We tested 128 Salmonella strains, representing all serotypes from A to Z and 51 to 66 as well as 50 non-Salmonella strains for inclusivity/exclusivity. Performance of the VIP using the new enrichment protocol was compared with the U.S. Department of Agriculture (USDA) Microbiology Laboratory Guidebook reference culture procedure for the detection of Salmonella in ready-to-eat poultry, roast beef, and chicken carcass rinsate. VIP performance was also compared with the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) for the detection of Salmonella from raw spinach, raw almonds, raw pasta, and environmental surfaces (stainless steel, rubber, and plastic). Results: The VIP detected all 128 of Salmonella strains and none of the 50 non-Salmonella strains. There was no statistically significant difference in the numbers of positive results with VIP Gold for Salmonella protocol compared with appropriate USDA-Food Safety and Inspection Service or FDA-BAM reference methods for any of these matrixes. Conclusions: This new enrichment protocol has met all the requirements to be approved as a Performance Tested MethodSM. Highlights: The new enrichment protocol will improve the time-to-results and allow quicker decisions about the contamination of food products.

Comparative Validation Study to Demonstrate the Equivalence of an Alternate Next-Day Enrichment Protocol for the TRANSIA® PLATE Salmonella Gold Method to Culture Methods for the Detection of Salmonella in Selected Foods and Environmental Surfaces

2019 ◽  
Vol 102 (3) ◽  
pp. 828-841
Author(s):  
David E Kerr ◽  
George Shen ◽  
Andrew H Lienau ◽  
Ta Deng ◽  
Mandeep Kaur ◽  
...  
Keyword(s):  
Enterohemorrhagic Escherichia Coli ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Method Performance ◽  
Environmental Surfaces ◽  
Chicken Carcass ◽  
Significant Difference ◽  
Enrichment Protocol ◽  
Inspection Service ◽  
Comparative Validation

Abstract TRANSIA® PLATE Salmonella Gold is an ELISA that was validated by Association Française de Normalisation (AFNOR) in 2001 and as a Performance Tested MethodSM (PTM) by AOAC in 2006 (PTM No. 010602) as a two-step enrichment protocol requiring 48 h. A simple next-day enrichment protocol using modified Enterohemorrhagic Escherichia coli media was developed for the TRANSIA PLATE Salmonella Gold to improve the time-to-results and laboratory work flow. We tested 128 Salmonella strains, representing all serotypes from A though Z and 51–66. TRANSIA PLATE Salmonella Gold detected all 128 of these strains. None of the 50 non-Salmonella strains were detected by TRANSIA PLATE Salmonella Gold. Performance of TRANSIA PLATE Salmonella Gold using the new enrichment protocol was compared with U.S. Department of Agriculture Microbiology Laboratory Guidebook reference culture procedure for the detection of Salmonella in ready-to-eat poultry, ready-to-eat beef, and chicken carcass rinsate. In addition, TRANSIA PLATE Salmonella Gold performance was compared with U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) for the detection of Salmonella from raw spinach, raw almonds, raw pasta, and environmental surfaces (stainless steel, rubber, and plastic). There was no statistically significant difference in the numbers of positive results TRANSIA PLATE Salmonella Gold protocol compared with the appropriate U.S. Department of Agriculture Food Safety and Inspection Service or FDA-BAM reference methods for any of these matrixes. Robustness testing demonstrated that the introduction of small changes in the normal assay parameters had no impact on the method performance. This new enrichment protocol has been approved as a Third Level modification to Performance Tested Method 010602.

Immunomagnetic Separation and Visual Fluorescence Detection of Salmonella spp., Using AOAC Approved SASTM Molecular Tests

2014 ◽  
Vol 97 (4) ◽  
pp. 1067-1072
Author(s):  
Chandra Bapanpally ◽  
Gayatri Maganty ◽  
Shah Khan ◽  
Akif Kasra ◽  
Amit Morey
Keyword(s):  
Reference Method ◽  
Current Method ◽  
Immunomagnetic Separation ◽  
Salmonella Spp ◽  
Test Portion ◽  
Molecular Tests ◽  
Modified Method ◽  
Reference Methods ◽  
Chicken Carcass ◽  
The U.S

Abstract The SASTM Molecular Tests method for detection of Salmonella spp. in various food matrixes has been certified by the AOAC Research Institute and designated Performance Tested MethodSM No. 021202. The current method modification includes the optional immunomagnetic separation (IMS) to enrich the bacteria as well as optional visual fluorescence readout without the use of a turbidimeter. The modifications were validated against the U.S. Department of Agriculture/Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG) and the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) reference methods. Food matrixes (chicken carcass rinse, beef trim, and spinach) were inoculated with low levels of Salmonella spp. (0.2–2 CFU/test portion) to generate fractional positives (5–15) in 20 inoculated samples. Samples were enriched with SAS Enrichment medium and incubated at 42 ± 1°C. Enrichments were tested directly and subjected to anti-Salmonella IMS prior to the SAS Molecular Tests. Results were determined via visual fluorescence and via turbidity using a turbidimeter. All replicates were confirmed using the MLG or BAM reference method procedures, regardless of presumptive result. The SAS Molecular Tests Salmonella Detection modified methods were determined to be equivalent to the reference methods for the detection of Salmonella in chicken carcass rinse, beef trim, and fresh spinach. The inclusion of IMS in the modified method improved the detection rate of Salmonella in chicken carcass rinses and spinach. The optional use of visual fluorescent reagent and heat block either with IMS or without IMS produced results that were comparable to the results obtained from using a real-time turbidimeter.

Validation of Predictive Mathematical Models Describing the Growth of Escherichia coli O157:H7 in Raw Ground Beef

1996 ◽  
Vol 59 (12) ◽  
pp. 1331-1335 ◽  
Author(s):  
ISABEL WALLS ◽  
VIRGINIA N. SCOTT
Keyword(s):  
Escherichia Coli ◽  
Predictive Models ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Growth Data ◽  
Department Of Agriculture ◽  
Gompertz Equation ◽  
Temperature Requirements ◽  
Inspection Service ◽  
The U.S

The growth of Escherichia coli O157:H7 in raw ground beef was investigated at 12°C, 20°C, and 35°C at pH 5.7 (unadjusted) and adjusted to pH 6.3 to 6.4. These growth data were fitted to the Gompertz equation and the resulting growth kinetics were compared with predictions from the U.S. Department of Agriculture Pathogen Modeling Program. Close agreement with the model was obtained at pH 5.7, but at pH 6.4, growth was more rapid than predicted. The U.S. Department of Agriculture Food Safety and Inspection Service has used this predictive model for developing proposed regulations on time-temperature requirements for carcass cooling. As there may be considerable differences in the microenvironment of raw ground beef and a beef carcass, the validity of using predictive models for estimating growth rates on a carcass should be determined by performing growth studies on carcass surfaces.

Validation of the ANSR®E. coli O157:H7 Method for Detection of E. coli O157:H7

2016 ◽  
Vol 99 (3) ◽  
pp. 705-716 ◽  
Author(s):  
Ryan Viator ◽  
Susan Alles ◽  
Quynh-Nhi Le ◽  
Edan Hosking ◽  
Evan Meister ◽  
...  
Keyword(s):  
Target Genes ◽  
Ground Beef ◽  
Reference Method ◽  
Single Step ◽  
Fluorescence Detector ◽  
Method Performance ◽  
E Coli ◽  
Reference Methods ◽  
Inspection Service ◽  
The U.S

Abstract A performance validation of the ANSR® for E. coli O157:H7 method was conducted in selected food matrixes. This assay uses selective nicking enzyme amplification technology to amplify target genes. Samples are enriched for 12–24 h and then lysed. The assay is completed within 40 min using real-time detection in a combination incubator/fluorescence detector and software. When 44 distinct strains of Escherichia coli O157:H7 and 6 strains of E. coli O157:NM were tested for inclusivity, all 50 strains produced positive results. In exclusivity testing, 57 strains representing 33 species of closely related Gram-negative bacteria belonging to the Enterobacteriaceae family, including 11 non-H7 O157 strains and shiga toxin-producing E. coli other than O157:H7, were evaluated. All 57 nontarget strains generated negative ANSR assay results. Using 80% lean ground beef and beef trim (approximately 20% fat), ANSR method performance was compared to the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference culture procedure. ANSR performance with baby spinach and sprout irrigation water was measured against the U.S. Food and Drug Administration Bacteriological Analytical Manual reference method. ANSR method performance was not statistically different to that of the reference methods using two different enrichment options. For ground beef and beef trim, the standard enrichment in modified Tryptone Soya Broth can be analyzed using the ANSR assay with a 1:10 dilution of the enrichment in phosphate-buffered saline and produces equivalent results to the reference method. Additionally, in most matrixes tested (exception is spinach which required 24 h enrichment) the assay offers great efficiency and flexibility over the reference method with a 12–24 h single-step enrichment. Equivalent results were observed at both time points (12 and 24 h) to reference methods. Small changes to the assay parameters minimally affected ANSR method performance. Finally, accelerated stability results from three independently manufactured lots support a shelf-life of 6 months when stored at 4°C.

Evaluation of Modification of the 3M™ Molecular Detection Assay (MDA) Salmonella Method (2013.09) for the Detection of Salmonella in Selected Foods: Collaborative Study

2014 ◽  
Vol 97 (5) ◽  
pp. 1329-1342 ◽  
Author(s):  
Patrick Bird ◽  
Kiel Fisher ◽  
Megan Boyle ◽  
Travis Huffman ◽  
M Joseph Benzinger ◽  
...  
Keyword(s):  
Confidence Intervals ◽  
Molecular Detection ◽  
Collaborative Study ◽  
Ground Beef ◽  
Inoculum Level ◽  
Test Portion ◽  
Low Level ◽  
Dog Food ◽  
Detection Assay ◽  
The U.S

Abstract The 3M™ Molecular Detection Assay (MDA) Salmonella utilizes isothermal amplification of nucleic acid sequences with high specificity, efficiency, rapidity and bioluminescence to detect amplification of Salmonella spp. in food, food-related, and environmental samples after enrichment. A method modification and matrix extension study of the previously approved AOAC Official MethodSM 2013.09 was conducted, and approval of the modification was received on March 20, 2014. Using an unpaired study design in a multilaboratory collaborative study, the 3M MDA Salmonella method was compared to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.05 (2011), Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Catfish Products for raw ground beef and the U.S. Food and Drug Administration (FDA)/Bacteriological Analytical Manual (BAM) Chapter 5, Salmonella reference method for wet dog food following the current AOAC guidelines. A total of 20 laboratories participated. For the 3M MDA Salmonella method, raw ground beef was analyzed using 25 g test portions, and wet dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. Each matrix 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, 1512 unpaired replicate samples were analyzed. Statistical analysis was conducted according to the probability of detection (POD). For the low-level raw ground beef test portions, the following dLPOD (difference between the LPODs of the reference and candidate method) values with 95% confidence intervals were obtained: –0.01 (–0.14, +0.12). For the low-level wet dog food test portions, the following dLPOD with 95% confidence intervals were obtained: –0.04 (–0.16, +0.09). No significant differences were observed in the number of positive samples detected by the 3M MDA Salmonella method versus either the USDA/FSIS-MLG or FDA/BAM methods.

Contamination Revealed by Indicator Microorganism Levels during Veal Processing

2016 ◽  
Vol 79 (8) ◽  
pp. 1341-1347 ◽  
Author(s):  
JOSEPH M. BOSILEVAC ◽  
RONG WANG ◽  
BRANDON E. LUEDTKE ◽  
TOMMY L. WHEELER ◽  
MOHAMMAD KOOHMARAIE
Keyword(s):  
Escherichia Coli ◽  
Plate Count ◽  
Department Of Agriculture ◽  
E Coli ◽  
Veal Calves ◽  
Indicator Microorganism ◽  
Analysis Of Results ◽  
Aerobic Plate Count ◽  
Inspection Service ◽  
The U.S

ABSTRACT During site visits of veal processors, the U.S. Department of Agriculture, Food Safety Inspection Service (FSIS) has reported processing deficiencies that likely contribute to increased levels of veal contamination. Here, we report the results of measuring aerobic plate count bacteria (APC), Enterobacteriaceae, coliforms (CF), and Escherichia coli during eight sample collections at five veal processors to assess contamination during the harvest of bob veal and formula-fed veal before (n = 5 plants) and after (n = 3 plants) changes to interventions and processing practices. Hides of veal calves at each plant had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 6.02 to 8.07, 2.95 to 5.24, 3.28 to 5.83, and 3.08 to 5.59, respectively. Preintervention carcasses had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 3.08 to 5.22, 1.16 to 3.47, 0.21 to 3.06, and −0.07 to 3.10, respectively, before and 2.72 to 4.50, 0.99 to 2.76, 0.69 to 2.26, and 0.33 to 2.12, respectively, after changes were made to improve sanitary dressing procedures. Final veal carcasses had mean log CFU/100 cm2 APC, Enterobacteriaceae, CF, and E. coli of 0.36 to 2.84, −0.21 to 1.59, −0.23 to 1.59, and −0.38 to 1.45 before and 0.44 to 2.64, −0.16 to 1.33, −0.42 to 1.20, and −0.48 to 1.09 after changes were made to improve carcass-directed interventions. Whereas the improved dressing procedures resulted in improved carcass cleanliness, the changes to carcass-directed interventions were less successful, and veal processors are urged to use techniques that ensure uniform and consistent delivery of antimicrobials to carcasses. Analysis of results comparing bob veal to formula-fed veal found bob veal hides, preintervention carcasses, and final carcasses to have increased (P < 0.05) APC, Enterobacteriaceae, CF, and E. coli (with the exception of hide Enterobacteriaceae; P > 0.05) relative to formula fed veal. When both veal categories were harvested at the same plant on the same day, similar results were observed. Since identification by FSIS, the control of contamination during veal processing has started to improve, but challenges still persist.

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