Vivione Bioscience RAPID-B®E. coli O157 Test Kit and non-O157 STEC Test Kit Evaluation

2015 ◽  
Vol 98 (2) ◽  
pp. 371-378 ◽  
Author(s):  
Melinda Miller ◽  
Shawn Ramsaroop ◽  
Chris Lopez ◽  
Bharath Brahmanda
Keyword(s):  
High Performance ◽  
Diagnostic System ◽  
Brain Heart Infusion ◽  
Reference Method ◽  
Test Methods ◽  
Cell Surface Antigens ◽  
Test Portion ◽  
E Coli ◽  
Test Kit ◽  
B Test

Abstract RAPID-B® is a high performance, integrated microbiology/infectious disease diagnostic system. The system uses hardware and software that are specifically designed for optimal detection using custom, immuno-based reagents designed to react to cell surface antigens of the target bacteria. The Vivione Bioscience RAPID-B Escherichia coli O157 and non-O157 Shiga toxin-producing E. coli (STEC) kits were validated alongside the U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS), Microbiology Laboratory Guidebook (MLG) 5.07 (for E. coli O157) and FSIS MLG 5B.04 (fornon-O157 STEC) reference methods for the detection of E. coli O157 and STEC. The matrixes, ground beef and beef trim, were inoculated with appropriate CFU/test portion of E. coli O157 and STEC so as to generate fractional positives results, 5 to 15 positives out of 20 inoculated samples. Samples were enriched in prewarmed Brain Heart Infusion broth at 42 ± 1°C for 6.5–7.5 h or 8.5–9.5 h depending on thesample size. All samples were confirmed using the MLG reference method, regardless of initial screen result. The RAPID-B test methods were statistically equivalent to the reference method for the detection ofE. coli O157 and STEC in all testedsamples. Inclusivity and exclusivity testing of the RAPID-B methods showed 100% specificity for both kits. Finally, the RAPID-B test methods were shown to be robust when variations were applied to enrichment time, broth temperature, and vortexing time.

scholarly journals Validation of iQ-Check E. coli O157:H7 Real-Time PCR Test Kit for Detection of Escherichia coli O157:H7 in Selected Foods

2009 ◽  
Vol 92 (4) ◽  
pp. 1095-1104 ◽  
Author(s):  
Wendy F Lauer ◽  
Sylvie Tymciu ◽  
Caroline D Sidi ◽  
Pierre Sonigo
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Ground Beef ◽  
Reference Method ◽  
Target Sequence ◽  
Apple Cider ◽  
E Coli ◽  
Test Kit ◽  
Significant Difference ◽  
The U.S

Abstract iQ-Check E. coli O157:H7 (Bio-Rad Laboratories, Hercules, CA) is a real-time PCR kit for detection of E. coli O157:H7 from selected foods. Specific fluorescent oligonucleotide probes are used to detect target DNA during the amplification, by hybridizing to the amplicons. These fluorescent probes are linked to a fluorophore which fluoresces only when hybridized to the target sequence. Three foods (ground beef, apple cider, fresh spinach) were selected to compare the performance of iQ-Check E. coli O157:H7 to the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG) reference method for ground beef and the U.S. Food and Drug Administration/Bacteriological Analytical Manual reference method for apple cider and fresh spinach. Three protocols were tested in this study: a shortened 8 h primary enrichment in buffered peptone water (BPW), a 24 h enrichment in BPW, and an enrichment in appropriate reference method enrichment broth. The iQ-Check E. coli O157:H7 method was able to identify more true/confirmed positive samples than the reference method. Inclusivity and exclusivity rates of the method were 100. iQ-Check E. coli O157:H7 performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no significant difference observed in performance over the shelf life of the kit.

scholarly journals Modification of the BAX® System PCR Assay for Detecting Salmonella in Beef, Produce, and Soy Protein Isolate

2011 ◽  
Vol 94 (1) ◽  
pp. 172-178 ◽  
Author(s):  
Linda X Peng ◽  
Morgan Wallace ◽  
Bridget Andaloro ◽  
Dawn Fallon ◽  
Lois Fleck ◽  
...  
Keyword(s):  
Soy Protein ◽  
Soy Protein Isolate ◽  
Method Comparison ◽  
Brain Heart Infusion ◽  
Reference Method ◽  
Culture Method ◽  
Protein Isolate ◽  
Pcr Assay ◽  
E Coli ◽  
The U.S

Abstract The BAX® System PCR assay for Salmonella detection in foods was previously validated as AOAC Research Institute (RI) Performance Tested MethodSM (PTM) 100201. New studies were conducted on beef and produce using the same media and protocol currently approved for the BAX System PCR assay for E. coli O157:H7 multiplex (MP). Additionally, soy protein isolate was tested for matrix extension using the U.S. Food and Drug Administration-Bacteriological Analytical Manual (FDA-BAM) enrichment protocols. The studies compared the BAX System method to the U.S. Department of Agriculture culture method for detecting Salmonella in beef and the FDA-BAM culture method for detecting Salmonella in produce and soy protein isolate. Method comparison studies on low-level inoculates showed that the BAX System assay for Salmonella performed as well as or better than the reference method for detecting Salmonella in beef and produce in 8–24 h enrichment when the BAX System E. coli O157:H7 MP media was used, and soy protein isolate in 20 h enrichment with lactose broth followed by 3 h regrowth in brain heart infusion broth. An inclusivity panel of 104 Salmonella strains with diverse serotypes was tested by the BAX System using the proprietary BAX System media and returned all positive results. Ruggedness factors involved in the enrichment phase were also evaluated by testing outside the specified parameters, and none of the factors examined affected the performance of the assay.

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.

Method Modification of the Legipid®Legionella Fast Detection Test Kit

2014 ◽  
Vol 97 (5) ◽  
pp. 1403-1409 ◽  
Author(s):  
Guillermo Rodríguez Albalat ◽  
Begoña Bedrina Broch ◽  
Marisa Jiménez Bono
Keyword(s):  
Legionella Pneumophila ◽  
Reference Method ◽  
Culture Method ◽  
Test Method ◽  
Magnetic Microspheres ◽  
Test Portion ◽  
Relative Level ◽  
Fast Detection ◽  
Test Kit ◽  
Optical Reader

Abstract Legipid®Legionella Fast Detection is a test based on combined magnetic immunocapture and enzyme-immunoassay (CEIA) for the detection of Legionella in water. The test is based on the use of anti-Legionella antibodies immobilized on magnetic microspheres. Target microorganism is preconcentrated by filtration. Immunomagnetic analysis is applied on these preconcentrated water samples in a final test portion of 9 mL. The test kit was certified by the AOAC Research Institute as Performance Tested MethodSM (PTM) No. 111101 in a PTM validation which certifies the performance claims of the test method in comparison to the ISO reference method 11731-1998 and the revision 11731-2004 “Water Quality: Detection and Enumeration of Legionella pneumophila” in potable water, industrial water, and waste water. The modification of this test kit has been approved. The modification includes increasing the target analyte from L. pneumophila to Legionella species and adding an optical reader to the test method. In this study, 71 strains of Legionella spp. other than L. pneumophila were tested to determine its reactivity with the kit based on CEIA. All the strains of Legionella spp. tested by the CEIA test were confirmed positive by reference standard method ISO 11731. This test (PTM 111101) has been modified to include a final optical reading. A methods comparison study was conducted to demonstrate the equivalence of this modification to the reference culture method. Two water matrixes were analyzed. Results show no statistically detectable difference between the test method and the reference culture method for the enumeration of Legionella spp. The relative level of detection was 93 CFU/volume examined (LOD50). For optical reading, the LOD was 40 CFU/volume examined and the LOQ was 60 CFU/volume examined. Results showed that the test Legipid Legionella Fast Detection is equivalent to the reference culture method for the enumeration of Legionella spp.

scholarly journals Evaluation of the GENE-UP ®E. coli O157:H7 Method for the Detection of E. coli O157:H7 in Select Foods: Collaborative Study, 2019.03

2020 ◽  
Vol 103 (5) ◽  
pp. 1338-1347
Author(s):  
Ronald Johnson ◽  
John Mills ◽  
Jean-Louis Pittet ◽  
Maryse Rannou ◽  
Patrick Bird ◽  
...  
Keyword(s):  
Confidence Interval ◽  
Collaborative Study ◽  
Control Level ◽  
Reference Method ◽  
Probability Of Detection ◽  
Contamination Level ◽  
Test Portion ◽  
E Coli ◽  
Significant Difference ◽  
Contamination Levels

Abstract Background The GENE-UP®E. coli O157:H7 2 (ECO 2) assay (Performance Tested MethodSM 121805) incorporates Fluorescence Resonance Energy Transfer hybridization probes into its proprietary PCR technology for the rapid detection of E. coli O157:H7 in select foods. 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 E. coli O157:H7 in select foods. Method The GENE-UP® method was evaluated in a multi-laboratory study as part of the MicroVal validation process using unpaired test portions for one food matrix, raw milk cheese (Comté, 34% fat, 0.8% salt). The candidate method was compared to the ISO 16654:2001 reference method. Fourteen participants from 13 laboratories throughout the European Union participated. Three levels of contamination were evaluated: a non-inoculated control level (0 colony-forming units (CFU)/test portion), a low contamination level (∼5 CFU/test portion), and a high contamination level (∼10 CFU/test portion). Data from that study were analyzed according to the Probability of Detection (POD) statistical model as presented in the AOAC validation guidelines. The difference in laboratory POD (dLPODC) values with 95% confidence interval across collaborators was calculated for each level between the candidate and reference method results, and between the candidate presumptive and confirmed results. Results The dLPODC values with 95% confidence interval were; 0.00 (–0.04, 0.04), 0.27 (0.04, 0.49), and 0.17 (0.01, 0.33) for the non-inoculated, low and high contamination levels respectively. Conclusions The dLPODC results indicate a significant difference between the candidate method and the reference method for both the low and high contamination levels, with the candidate method producing higher recovery of the target organism at both levels. Highlights The GENE-UP E. coli O157:H7 assay provides industry with a rapid, accurate detection method for E. coli O157:H7 in a broad range of foods.

Evaluation of the GENE-UP® EHEC Detection Method for the Detection of Enterohemorrhagic E. coli in Select Foods: Collaborative Study: First Action Method 2020.06

2021 ◽  
Author(s):  
Ronald Johnson ◽  
John Mills ◽  
Jean-Louis Pittet ◽  
Maryse Rannou ◽  
Patrick Bird
Keyword(s):  
Detection Method ◽  
Collaborative Study ◽  
False Negative ◽  
Control Level ◽  
Reference Method ◽  
Probability Of Detection ◽  
Contamination Level ◽  
Official Method ◽  
Test Portion ◽  
E Coli

Abstract Background The GENE-UP® EHEC assay (Performance Tested MethodSM 121806) is a real-time PCR molecular detection method that utilizes Fluorescence Resonance Energy Transfer proprietary hybridization probes for the rapid detection of Enterohemorrhagic E. coli (EHEC) in select foods. Objective The purpose of this validation was to evaluate the method’s interlaboratory performance and submit the results to AOAC INTERNATIONAL for adoption as First Action Official Method of AnalysisSM for the detection of EHEC in select foods. Method The GENE-UP® method was evaluated in a multi-laboratory study as part of the MicroVal VALIDATION certification process using unpaired test portions for one food matrix, raw ground beef (85% lean). Collaborators evaluated the candidate method using either an automated or manual lysis procedure. The candidate method was compared to the ISO/TS 13136:2012 method. Data from 17 participants from 15 laboratories throughout the European Union was evaluated. Three levels of contamination were evaluated: a non-inoculated control level (0 CFU/test portion), a low contamination level (∼1 CFU/test portion) and a high contamination level (∼10 CFU/test portion). Data from the study were analyzed according to the probability of detection (POD) statistical model. Results The dLPODC values with 95% confidence interval between the candidate and reference method results were; –0.01 (–0.04, 0.02), 0.23 (0.07, 0.39) and 0.06 (0.01, 0.12) for the non-inoculated, low and high contamination levels, respectively. Conclusion For the candidate method, values obtained for repeatability and reproducibility were similar to the reference method and indicated minimal variation between samples or between laboratories. No discrepant results (false positive or false negative) were observed for each contamination. A statistical difference was calculated between the candidate and reference method at the low and high inoculation levels, with the candidate method detecting a higher number of positive samples indicating a higher sensitivity than the reference method. No differences in the recovery of the target analyte were observed between the manual and automated lysis procedures. Highlights The GENE-UP EHEC Detection Method provides end users a rapid, easy-to-use workflow for the detection of EHEC in food matrices.

Biodecomposition of Phenanthrene and Pyrene by a Genetically Engineered Escherichia coli

2020 ◽  
Vol 14 (2) ◽  
pp. 121-133 ◽  
Author(s):  
Maryam Ahankoub ◽  
Gashtasb Mardani ◽  
Payam Ghasemi-Dehkordi ◽  
Ameneh Mehri-Ghahfarrokhi ◽  
Abbas Doosti ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Performance ◽  
Human Cancer ◽  
Genetically Engineered ◽  
Hazardous Substances ◽  
E Coli ◽  
Spiked Soil ◽  
Engineered Escherichia Coli ◽  
Genetically Manipulated ◽  
Hplc Measurement

Background: Genetically engineered microorganisms (GEMs) can be used for bioremediation of the biological pollutants into nonhazardous or less-hazardous substances, at lower cost. Polycyclic aromatic hydrocarbons (PAHs) are one of these contaminants that associated with a risk of human cancer development. Genetically engineered E. coli that encoded catechol 2,3- dioxygenase (C230) was created and investigated its ability to biodecomposition of phenanthrene and pyrene in spiked soil using high-performance liquid chromatography (HPLC) measurement. We revised patents documents relating to the use of GEMs for bioremediation. This approach have already been done in others studies although using other genes codifying for same catechol degradation approach. Objective: In this study, we investigated biodecomposition of phenanthrene and pyrene by a genetically engineered Escherichia coli. Methods: Briefly, following the cloning of C230 gene (nahH) into pUC18 vector and transformation into E. coli Top10F, the complementary tests, including catalase, oxidase and PCR were used as on isolated bacteria from spiked soil. Results: The results of HPLC measurement showed that in spiked soil containing engineered E. coli, biodegradation of phenanthrene and pyrene comparing to autoclaved soil that inoculated by wild type of E. coli and normal soil group with natural microbial flora, were statistically significant (p<0.05). Moreover, catalase test was positive while the oxidase tests were negative. Conclusion: These findings indicated that genetically manipulated E. coli can provide an effective clean-up process on PAH compounds and it is useful for bioremediation of environmental pollution with petrochemical products.

scholarly journals Comparison of the ColiPlate™ Kit with Two Common E. coli Enumeration Methods for Water

Water ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 1804
Author(s):  
Cassi J. Gibson ◽  
Abraham K. Maritim ◽  
Jason W. Marion
Keyword(s):  
High Risk ◽  
Natural Water ◽  
Membrane Filtration ◽  
Fecal Contamination ◽  
Reference Method ◽  
World Health ◽  
Risk Levels ◽  
E Coli ◽  
Field Samples ◽  
Very High

Quantitatively assessing fecal indicator bacteria in drinking water from limited resource settings (e.g., disasters, remote areas) can inform public health strategies for reducing waterborne illnesses. This study aimed to compare two common approaches for quantifying Escherichia coli (E. coli) density in natural water versus the ColiPlate™ kit approach. For comparing methods, 41 field samples from natural water sources in Kentucky (USA) were collected. E. coli densities were then determined by (1) membrane filtration in conjunction with modified membrane-thermotolerant E. coli (mTEC) agar, (2) Idexx Quanti-Tray® 2000 with the Colilert® substrate, and (3) the Bluewater Biosciences ColiPlate kit. Significant correlations were observed between E. coli density data for all three methods (p < 0.001). Paired t-test results showed no difference in E. coli densities determined by all the methods (p > 0.05). Upon assigning modified mTEC as the reference method for determining the World Health Organization-assigned “very high-risk” levels of fecal contamination (> 100 E. coli CFU/100 mL), both ColiPlate and Colilert exhibited excellent discrimination for screening very high-risk levels according to the area under the receiver operating characteristic curve (~89%). These data suggest ColiPlate continues to be an effective monitoring tool for quantifying E. coli density and characterizing fecal contamination risks from water.

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