scholarly journals In-House Validation Study of the DuPont Qualicon BAX System Q7 Instrument with the BAX System PCR Assay for Salmonella (Modification of AOAC Official MethodSM 2003.09 and AOAC Research Institute Performance-Tested MethodSM 100201)

2009 ◽  
Vol 92 (3) ◽  
pp. 989-994 ◽  
Author(s):  
George Tice ◽  
Bridget Andaloro ◽  
H Kirk White ◽  
Lance Bolton ◽  
Siqun Wang ◽  
...  
Keyword(s):  
Validation Study ◽  
Reference Method ◽  
Pcr Assay ◽  
Paired Samples ◽  
Reference Methods ◽  
Comparable Performance ◽  
Aoac Official ◽  
Inspection Service ◽  
Positive Results ◽  
The U.S

Abstract In 2006, DuPont Qualicon introduced the BAX<sup/> system Q7 instrument for use with its assays. To demonstrate the equivalence of the new and old instruments, a validation study was conducted using the BAX system PCR Assay for Salmonella, AOAC Official MethodSM 2003.09, on three food types. The foods were simultaneously analyzed with the BAX system Q7 instrument and either the U.S. Food and Drug Administration Bacteriological Analytical Manual or the U.S. Department of AgricultureFood Safety and Inspection Service Microbiology Laboratory Guidebook reference method for detecting Salmonella. Comparable performance between the BAX system and the reference methods was observed. Of the 75 paired samples analyzed, 39 samples were positive by both the BAX system and reference methods, and 36 samples were negative by both the BAX system and reference methods, demonstrating 100 correlation. Inclusivity and exclusivity for the BAX system Q7 instrument were also established by testing 50 Salmonella strains and 20 non-Salmonella isolates. All Salmonella strains returned positive results, and all non-Salmonella isolates returned a negative response.

scholarly journals Modification of the BAX®Salmonella Test Kit to Include a Hot Start Functionality (Modification of AOAC Official MethodSM 2003.09)

2011 ◽  
Vol 94 (5) ◽  
pp. 1490-1505
Author(s):  
F Morgan Wallace ◽  
Deana DiCosimo ◽  
Andrew Farnum ◽  
George Tice ◽  
Bridget Andaloro ◽  
...  
Keyword(s):  
Reference Method ◽  
Department Of Agriculture ◽  
Reference Methods ◽  
Hot Start ◽  
System Method ◽  
Test Kit ◽  
Aoac Official ◽  
Inspection Service ◽  
Identical Performance ◽  
The U.S

Abstract In 2010, the BAX® System PCR assay for Salmonella was modified to include a hot start functionality designed to keep the reaction enzyme inactive until PCR begins. To validate the assay's Official Methods of Analysis status to include this procedure modification, an evaluation was conducted on four food types that were simultaneously analyzed with the BAX System and either the U.S. Food and Drug Administration's Bacteriological Analytical Manual or the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method for detecting Salmonella. Identical performance between the BAX System method and the reference methods was observed. Additionally, lysates were analyzed using both the BAX System Classic and BAX System Q7 instruments with identical results using both platforms for all samples tested. Of the 100 samples analyzed, 34 samples were positive for both the BAX System and reference methods, and 66 samples were negative by both the BAX System and reference methods, demonstrating 100% correlation. No instrument platform variation was observed. Additional inclusivity and exclusivity testing using the modified test kit demonstrated the test kit to be 100% accurate in evaluation of test panels of 352 Salmonella strains and 46 non-Salmonella strains.

scholarly journals Validation Study of a Rapid ELISA for Detection of Deoxynivalenol in Wheat, Barley, Malted Barley, Corn, Oats, and Rice

2010 ◽  
Vol 93 (2) ◽  
pp. 600-610 ◽  
Author(s):  
Anthony Lupo ◽  
Chris Roebuck ◽  
Ken Settimo ◽  
Anna Quain ◽  
Justina Kennedy ◽  
...  
Keyword(s):  
Rapid Method ◽  
Electron Capture ◽  
Validation Study ◽  
Reference Method ◽  
Elisa Test ◽  
Electron Capture Detection ◽  
Reference Methods ◽  
Test Kit ◽  
Aoac Official ◽  
Research Institute

Abstract Neogen Corp. developed the Veratox DON test kit for the detection of deoxynivalenol (DON). The purpose of this study was to validate the method under the requirements of the AOAC Research Institute Performance Tested MethodsSM (PTM) program. There are two AOAC Official MethodsSM for DON detection: 986.17 and 986.18, the first of which is a TLC method and the second a GC method. A rapid method (PTM 000701) has also been performance tested by the AOAC Research Institute. One of the most widely used reference methods; however, is a GC method with electron capture detection that is referred to as the reference method in this paper. Although considered the reference method, the GC procedure is complicated and requires the investment of both expensive equipment and a highly skilled technician. A rapid (e.g., ELISA) test kit needs to be validated by the AOAC Research Institute. The Veratox 2/3 method is highly reproducible with average CV values <10, and is very accurate, showing >97 correlation to reference methods.

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.

scholarly journals USDA FSIS and FDA BAM Culture Methods BBL CHROMagar Salmonella Prepared Plated and Difco Dehydrated Culture Media

2009 ◽  
Vol 92 (2) ◽  
pp. 459-470 ◽  
Author(s):  
Vicki Ritter ◽  
Nancy Dick
Keyword(s):  
Culture Media ◽  
False Positive Rate ◽  
Ground Beef ◽  
Reference Method ◽  
Chi Square ◽  
Shell Eggs ◽  
Reference Methods ◽  
Raw Fish ◽  
Chi Square Analysis ◽  
The U.S

Abstract BBL and Difco CHROMagar Salmonella (CS) was evaluated internally and externally for the recovery of Salmonella in raw chicken, raw ground beef, raw fish, lettuce, and shell eggs. The raw chicken and ground beef were processed according to the U.S. Department of Agriculture, Food Safety and Inspection Service reference methods. The raw fish, lettuce, and shell eggs were processed according to the U.S. Food and Drug Administration, Bacteriological Analytical Manual procedures. Only raw chicken was found to be naturally contaminated with Salmonella; all other matrixeswere seededwith an appropriate dilution of organism to achieve fractionally positive results. Salmonella strains were permitted to equilibrate with the culture-negative matrixes for 48 h at 4C. Twenty 25 g samples of each food matrix plus 5 uninoculated samples were processed. CS prepared plates (CS PPM) and laboratory prepared plates from dehydrated culture media (CS DCM) were evaluated with the reference method media. A total of 16 positive cultures were obtained from the raw chicken samples, 17 in the raw ground beef, 18 in the raw fish and lettuce, and 11 in the shell eggs. A Chi-square analysis was performed on each of the food matrixes. BBL CS produced comparable results with the reference methods on all matrixes, resulting in a method agreement of 100 based on the Chi-square results. In testing known isolates the sensitivity and specificity was determined to be 94. Specificity improved to 98 when tetrathionate broth enrichment was used. A negative- and false-positive rate of 6 was found with known isolates. No false negatives were found in testing the food matrixes. The performance of the CS prepared plate and laboratory prepared plate was identical.

Evaluation of a Multiplex Real-Time PCR Method for Detecting Shiga Toxin–Producing Escherichia coli in Beef and Comparison to the U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook Method†

2014 ◽  
Vol 77 (2) ◽  
pp. 180-188 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
JAMIE L. WASILENKO ◽  
BRADLEY GARMAN ◽  
DANIEL R. DeMARCO ◽  
STEPHEN VARKEY ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Virulence Genes ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
Department Of Agriculture ◽  
E Coli ◽  
System Method ◽  
Inspection Service ◽  
The U.S

The “top-six” non-O157 Shiga toxin–producing Escherichia coli (STEC) serogroups (O26, O45, O103, O111, O121, and O145) most frequently associated with outbreaks and cases of foodborne illnesses have been declared as adulterants in beef by the U.S. Department of Agriculture Food Safety and Inspection Service (FSIS). Regulatory testing in beef began in June 2012. The purpose of this study was to evaluate the DuPont BAX System method for detecting these top six STEC strains and strains of E. coli O157:H7. For STEC, the BAX System real-time STEC suite was evaluated, including a screening assay for the stx and eae virulence genes and two panel assays to identify the target serogroups: panel 1 detects O26, O111, and O121, and panel 2 detects O45, O103, O145. For E. coli O157:H7, the BAX System real-time PCR assay for this specific serotype was used. Sensitivity of each assay for the PCR targets was ≥1.23 × 103 CFU/ml in pure culture. Each assay was 100% inclusive for the strains tested (20 to 50 per assay), and no cross-reactivity with closely related strains was observed in any of the assays. The performance of the BAX System methods was compared with that of the FSIS Microbiology Laboratory Guidebook (MLG) methods for detection of the top six STEC and E. coli O157:H7 strains in ground beef and beef trim. Generally, results of the BAX System method were similar to those of the MLG methods for detecting non-O157 STEC and E. coli O157:H7. Reducing or eliminating novobiocin in modified tryptic soy broth (mTSB) may improve the detection of STEC O111 strains; one beef trim sample inoculated with STEC O111 produced a negative result when enriched in mTSB with 8 mg/liter novobiocin but was positive when enriched in mTSB without novobiocin. The results of this study indicate the feasibility of deploying a panel of real-time PCR assay configurations for the detection and monitoring of the top six STEC and E. coli O157:H7 strains in beef. The approach could easily be adapted for additional multiplex assays should regulations expand to include other O serogroups or virulence genes.

Romer Labs RapidChek®Listeria monocytogenes Test System for the Detection of L. monocytogenes on Selected Foods and Environmental Surfaces

2018 ◽  
Vol 101 (5) ◽  
pp. 1490-1507
Author(s):  
Gregory Juck ◽  
Verapaz Gonzalez ◽  
Ann-Christine Olsson Allen ◽  
Meredith Sutzko ◽  
Kody Seward ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Reference Method ◽  
Test System ◽  
Probability Of Detection ◽  
Department Of Agriculture ◽  
Detection Analysis ◽  
Media System ◽  
Reference Methods ◽  
Environmental Surfaces ◽  
Inspection Service

Abstract The Romer Labs RapidChek®Listeria monocytogenes test system (Performance Tested Method 011805) was validated against the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (USDA-FSIS/MLG), U.S. Food and Drug Association Bacteriological Analytical Manual (FDA/BAM), and AOAC Official Methods of Analysis (AOAC/OMA) cultural reference methods for the detection of L. monocytogenes on selected foods including hot dogs, frozen cooked breaded chicken, frozen cooked shrimp, cured ham, and ice cream, and environmental surfaces including stainless steel and plastic in an unpaired study design. The RapidChek method uses a proprietary enrichment media system, a 44–48 h enrichment at 30 ± 1°C, and detects L. monocytogenes on an immunochromatographic lateral flow device within 10 min. Different L. monocytogenes strains were used to spike each of the matrixes. Samples were confirmed based on the reference method confirmations and an alternate confirmation method. A total of 140 low-level spiked samples were tested by the RapidChek method after enrichment for 44–48 h in parallel with the cultural reference method. There were 88 RapidChek presumptive positives. One of the presumptive positives was not confirmed culturally. Additionally, one of the culturally confirmed samples did not exhibit a presumptive positive. No difference between the alternate confirmation method and reference confirmation method was observed. The respective cultural reference methods (USDA-FSIS/MLG, FDA/BAM, and AOAC/OMA) produced a total of 63 confirmed positive results. Nonspiked samples from all foods were reported as negative for L. monocytogenes by all methods. Probability of detection analysis demonstrated no significant differences in the number of positive samples detected by the RapidChek method and the respective cultural reference method.

scholarly journals 1-2 Test

2009 ◽  
Vol 92 (6) ◽  
pp. 1846-1850 ◽  
Author(s):  
Philip Feldsine ◽  
Thomas Hammack
Keyword(s):  
Emergency Response ◽  
Peanut Butter ◽  
Reference Method ◽  
Statistical Analyses ◽  
Paired Samples ◽  
The Difference ◽  
Contamination Levels ◽  
The U.S ◽  
Research Institute

Abstract A study was conducted to determine the efficacy of detecting low contamination levels of Salmonella in peanut butter using the 1-2 Test. This study was conducted under the AOAC Research Institute Emergency Response Validation program. A set of samples was analyzed by the 1-2 Test and the U.S. Food and Drug Administration's Bacteriological Analytical Manual reference method. Among the 90 total samples and controls, 32 samples were positive by the 1-2 Test and the reference method. Statistical analyses of these paired samples indicated that for all levels analyzed, the difference between the 1-2 Test and reference method results were not statistically different at the 5 level.

scholarly journals DuPont Qualicon BAX® System Assay for Genus Listeria 24E

2011 ◽  
Vol 94 (3) ◽  
pp. 863-871
Author(s):  
F Morgan Wallace ◽  
Dawn Fallon ◽  
Daniel DeMarco ◽  
Stephen Varkey
Keyword(s):  
False Negative ◽  
Method Comparison ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Service Culture ◽  
Steel Surfaces ◽  
Consistent Performance ◽  
Inspection Service ◽  
Positive Results ◽  
The U.S

Abstract The new BAX® System PCR Assay for Genus Listeria 24E was evaluated for detecting Listeria spp. in frankfurters, spinach, cooked shrimp, queso fresco cheese, and on stainless steel surfaces with a single-stage enrichment in BAX System 24 Listeria Enrichment Broth (24 LEB). Method comparison studies performed on samples with low-level inoculates showed that the BAX System demonstrates a sensitivity equivalent or superior to the U.S. Food and Drug Administration's Bacteriological Analytical Manual and the U.S. Department of Agriculture-Food Safety and Inspection Service culture methods, but with a significantly shorter time to result. Tests to evaluate inclusivity and exclusivity returned no false-negative and no false-positive results on a diverse panel of isolates, and tests for lot-to-lot variability and tablet stability demonstrated consistent performance. Ruggedness studies determined that none of the factors examined, within the range of deviations from specified parameters examined, affect the performance of the assay.

Validation of the ANSR® Listeria monocytogenes Method for Detection of Listeria monocytogenes in Selected Food and Environmental Samples

2016 ◽  
Vol 99 (1) ◽  
pp. 112-123
Author(s):  
Oscar Caballero ◽  
Susan Alles ◽  
Quynh-Nhi Le ◽  
R Lucas Gray ◽  
Edan Hosking ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Storage Temperature ◽  
Reference Method ◽  
Single Step ◽  
Stainless Steel Surface ◽  
Method Performance ◽  
Inspection Service ◽  
Nicking Enzyme ◽  
Performance Results ◽  
The U.S

Abstract Work was conducted to validate performance of the ANSR® for Listeria monocytogenes method in selected food and environmental matrixes. This DNA-based assay involves amplification of nucleic acid via an isothermal reaction based on nicking enzyme amplification technology. Following single-step sample enrichment for 16–24 h for most matrixes, the assay is completed in 40 min using only simple instrumentation. When 50 distinct strains of L. monocytogenes were tested for inclusivity, 48 produced positive results, the exceptions being two strains confirmed by PCR to lack the assay target gene. Forty-seven nontarget strains (30 species), including multiple non-monocytogenes Listeria species as well as non-Listeria, Gram-positive bacteria, were tested, and all generated negative ANSR assay results. Performance of the ANSR method was compared with that of the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference culture procedure for detection of L. monocytogenes in hot dogs, pasteurized liquid egg, and sponge samples taken from an inoculated stainless steel surface. In addition, ANSR performance was measured against the U.S. Food and Drug Administration Bacteriological Analytical Manual reference method for detection of L. monocytogenes in Mexican-style cheese, cantaloupe, sprout irrigation water, and guacamole. With the single exception of pasteurized liquid egg at 16 h, ANSR method performance as quantified by the number of positives obtained was not statistically different from that of the reference methods. Robustness trials demonstrated that deliberate introduction of small deviations to the normal assay parameters did not affect ANSR method performance. Results of accelerated stability testing conducted using two manufactured lots of reagents predicts stability at the specified storage temperature of 4°C of more than 1 year.

Roka Listeria Detection Method Using Transcription Mediated Amplification to Detect Listeria Species in Select Foods and Surfaces

2012 ◽  
Vol 95 (6) ◽  
pp. 1672-1688 ◽  
Author(s):  
Hua Yang ◽  
Shannon Kaplan ◽  
Michael Reshatoff ◽  
Ernie Hu ◽  
Alexis Zukowski ◽  
...  
Keyword(s):  
Probability Of Detection ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Detection Analysis ◽  
Reference Methods ◽  
Smoked Salmon ◽  
Environmental Surfaces ◽  
Detection Assay ◽  
Inspection Service ◽  
The U.S

Abstract The Roka Listeria Detection Assay was compared to the reference culture methods for nine select foods and three select surfaces. The Roka method used Half-Fraser Broth for enrichment at 35 ± 2°C for 24–28 h. Comparison of Roka's method to reference methods requires an unpaired approach. Each method had a total of 545 samples inoculated with a Listeria strain. Each food and surface was inoculated with a different strain of Listeria at two different levels per method. For the dairy products (Brie cheese, whole milk, and ice cream), our method was compared to AOAC Official MethodSM993.12. For the ready-to-eat meats (deli chicken, cured ham, chicken salad, and hot dogs) and environmental surfaces (sealed concrete, stainless steel, and plastic), these samples were compared to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) method MLG 8.07. Cold-smoked salmon and romaine lettuce were compared to the U.S. Food and Drug Administration/Bacteriological Analytical Manual, Chapter 10 (FDA/BAM) method. Roka's method had 358 positives out of 545 total inoculated samples compared to 332 positive for the reference methods. Overall the probability of detection analysis of the results showed better or equivalent performance compared to the reference methods.

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