Elevated-Temperature, Colorimetric, Monoclonal, Enzyme-Linked Immunosorbent Assay for Rapid Screening of Salmonella in Foods: Collaborative Study

1994 ◽  
Vol 77 (2) ◽  
pp. 374-394 ◽  
Author(s):  
Karl F Eckner ◽  
Wendy A Dustman ◽  
Michael S Curiale ◽  
Russell S Flowers ◽  
Barbara J Robison
Keyword(s):  
Elevated Temperature ◽  
Immunosorbent Assay ◽  
Collaborative Study ◽  
False Negative ◽  
Culture Method ◽  
Soy Flour ◽  
Rapid Screening ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Milk Chocolate

Abstract A collaborative study was performed by 30 laboratories in 3 sets of trials to validate a modified colorimetric monoclonal enzyme-linked immunosorbent assay (ELISA) method for Salmonella detection. The modifications to the current methodology included incubation of enrichments and postenrichments at an elevated temperature, addition of novobiocin to the M-broth post-enrichment, and elimination of the centrifugation and agitation steps. Five artificially contaminated foods (nonfat dry milk, milk chocolate, dried egg, ground black pepper, and soy flour) and 1 naturally contaminated food (raw ground turkey) were analyzed. The artificially contaminated foods were inoculated with individual Salmonella serotypes at a high (10–50 cells/25 g) and low (1–5 cells/25 g) contamination level. Results from the modified ELISA method were compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method. In 2 of the food products, milk chocolate and pepper, a number of laboratories isolated Salmonella from uninoculated control samples, thus invalidating their data. As a result, there were too few laboratories remaining with valid data, and these foods were repeated. In the completed study, there were 11 false negative results obtained by the modified ELISA method, while there were 28 false negatives produced by the BAM/AOAC procedure. There were 11 ELISA positive assays which could not be confirmed by culture methods. Statistically, there were no differences between the modified, colorimetric, monoclonal ELISA and the reference culture method in all foods except raw turkey, where the ELISA method was more productive. The colorimetric monoclonal enzyme immunoassay (Salmonella-Tek) method for detecting Salmonella in all foods has been adopted first action by AOAC INTERNATIONAL.

scholarly journals LOCATE Enzyme-Linked Immunosorbent Assay for Detection of Salmonella in Food: Collaborative Study

1998 ◽  
Vol 81 (2) ◽  
pp. 419-437 ◽  
Author(s):  
Vidhya Gangar ◽  
Michael S Curiale ◽  
Armando D'onorio ◽  
Carol Donnelly ◽  
Paul Dunnigan ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Screening Method ◽  
Culture Method ◽  
Microtiter Plate ◽  
Black Pepper ◽  
Soy Flour ◽  
Food Type ◽  
Enzyme Linked Immunosorbent Assay ◽  
Data Sets ◽  
Whole Egg

abstract A collaborative study was performed in 27 laboratories to validate the enzyme-linked immunosorbent procedure LOCATE for rapid detection of Salmonella in foods. Results were read visually and with a microtiter plate reader. The LOCATE method was compared with the Bacteriological Analytical Manual (BAM)/AOAC INTERNATIONAL culture method for detecting Salmonella in 6 foods: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. Two foods—dried whole egg and black pepper—required repeat rounds because insufficient data sets were produced initially (AOAC INTERNATIONAL stipulates a minimum of 15 sets per food type). Each laboratory tested one or more of the 6 foods. A total of 1 439 samples were analyzed, and no significant differences (P <0.05) were observed between LOCATE with either visual or reader detection and BAM/AOAC INTERNATIONAL results. The LOCATE screening method with visual or reader detection is recommended for Official First Action Approval

Improved Hydrophobic Grid Membrane Filter Method, Using EF-18 Agar, for Detection of Salmonella in Foods: Collaborative Study

1990 ◽  
Vol 73 (5) ◽  
pp. 734-742
Author(s):  
Phyllis Entis
Keyword(s):  
Membrane Filter ◽  
Collaborative Study ◽  
False Negative ◽  
Screening Method ◽  
Culture Method ◽  
Soy Flour ◽  
Poultry Meat ◽  
Filter Method ◽  
Negative Results ◽  
False Negative Results

Abstract A collaborative study was carried out in 30 laboratories to validate Improvements to the official final action hydrophobic grid membrane filter (HGMF) screening method for Salmonella in foods, 985.42, by comparing the performance of the improved HGMF method against that of the AOAC/BAM conventional culture method. Six products were Included In the collaborative study: milk chocolate, raw deboned poultry meat, black pepper, soy flour, egg yolk powder, and nonfat dry milk. The raw deboned poultry meat was naturally contaminated with Salmonella, and the remaining 5 products were each Inoculated In advance with low levels of Individual Salmonella serotypes. The AOAC/BAM method produced 11 false negative results and the Improved HGMF method produced 18 false negative results. The improved HGMF Salmonella method has been approved Interim official first action for all foods to replace the HGMF official final action method, 985.42.

scholarly journals VIDAS Enzyme-Linked Fluorescent Immunoassay for Detection of Salmonella in Foods: Collaborative Study

1997 ◽  
Vol 80 (3) ◽  
pp. 491-504 ◽  
Author(s):  
Michael S Curiale ◽  
Vidhya Gangar ◽  
Carol Gravens ◽  
J R Agin ◽  
A Bound ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Culture Method ◽  
Black Pepper ◽  
Soy Flour ◽  
Milk Chocolate ◽  
Dry Milk ◽  
Nonfat Dry Milk ◽  
Whole Egg

Abstract The VIDAS SLM method for detection of Salmonella was compared with the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Twenty laboratories participated in the evaluation. Each laboratory tested one or more of 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. No significant differences (P< 0.05) were observed between the 2 methods. The 2 methods were in agreement for 99% of 1544 samples analyzed. Of the 20 samples out of agreement, 8 were VIDAS SLM positive and BAM/AOAC negative, and 12 were VIDAS SLM negative and BAM/AOAC positive. The VIDAS SLM method for detection of Salmonella in foods has been adopted first action by AOAC INTERNATIONAL.

scholarly journals Evaluation of VIDAS® Immuno-Concentration Salmonella/VIDAS Salmonella Immunoassay Method for Detection of Salmonella in Selected Foods: Collaborative Study

2002 ◽  
Vol 85 (3) ◽  
pp. 609-625 ◽  
Author(s):  
Wendy A Lepper ◽  
Ann M Schultz ◽  
Michael S Curiale ◽  
Ronald L Johnson ◽  
J Agin ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Culture Method ◽  
Black Pepper ◽  
Soy Flour ◽  
Milk Chocolate ◽  
Dry Milk ◽  
Nonfat Dry Milk ◽  
Whole Egg

Abstract The VIDAS Immuno-concentration Salmonella (ICS)/VIDAS Salmonella (SLM) immunoassay method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1266 of the 1440 samples. Of the 174 samples not in agreement, 69 were VIDAS ICS/SLM-positive and BAM/AOAC-negative and 105 were VIDAS ICS/SLM-negative and BAM/AOAC-positive.

Enzyme-Linked Immunosorbent Assay for Detection of Zearalenone in Corn, Wheat, and Pig Feed: Collaborative Study

1994 ◽  
Vol 77 (6) ◽  
pp. 1500-1508 ◽  
Author(s):  
Glenn A Bennett ◽  
Terry C Nelsen ◽  
Brjnton M Miller
Keyword(s):  
Immunosorbent Assay ◽  
Collaborative Study ◽  
Screening Method ◽  
The United States ◽  
False Positives ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
False Negatives ◽  
Relative Standard ◽  
Pig Feed

Abstract A direct competitive enzyme-linked immunosorbent assay (ELISA) screening method for zearalenone in corn, wheat, and feed at 500 ng/g was evaluated by 23 collaborators (22 laboratories) in an international collaborative study. Eighteen samples of spiked or naturally contaminated corn, wheat, and pig feed were prepared by the sponsoring laboratory and sent for testing with complete test kits to participating collaborators in Canada, Italy, Sweden, The Netherlands, and the United States. Test samples were extracted with methanolwater solution (70 + 30) by shaking on a wrist-action shaker for 3 min. A portion of the extract was mixed with an equal volume of zearalenone-enzyme conjugate, and the mixture was incubated with zearalenone-specific monoclonal antibodies coated onto microtiter wells. All test samples were assayed in duplicate. One of 52 (2%) blanks was reported positive. Thirty-nine of the 52 (75%) samples that were spiked at 500 ng/g were reported as positive. Forty-nine of the 51 (96%) samples with concentrations at or above 1000 ng/g were reported as positive. The overall incidence of false negatives was 6.0% and the incidence of false positives was 22.7% by the ELISA method. Only one (3.4%) false negative was reported for samples containing ≥800 ng/g. In the spectrophotometric method, 8 collaborators determined approximate levels of zearalenone in test samples from standard curves constructed from spiked extracts (0–3000 ng/g of each commodity tested). This method gave and overall incidence of false negatives of 5.7% and false positives of 17.8%. Average relative standard deviations, RSDr (repeatability) and RSDR (reproducibility), were 11.6 and 25.1% for spiked samples and 11.7 and 33.1% for naturally contaminated samples, respectively. Standard curves were constructed with each set of samples assayed. Comparison of absorbance values from these standard curves indicate the performance of reagents and antibody used in the assay. The ELISA method has been adopted first action by AOAC INTERNATIONAL as a screening method for zearalenone at ≥800 ng/g in corn, wheat, and pig feed.

Polyclonal Enzyme Immunoassay Method for Detection of Motile and Non-Motile Salmonella in Foods: Collaborative Study

1992 ◽  
Vol 75 (6) ◽  
pp. 1032-1044 ◽  
Author(s):  
Philip T Feldsine ◽  
Maria T Falbo-Nelson ◽  
David L Hustead
Keyword(s):  
Enzyme Immunoassay ◽  
Polyclonal Antibodies ◽  
Collaborative Study ◽  
False Negative ◽  
Culture Method ◽  
Black Pepper ◽  
Soy Flour ◽  
Agreement Rate ◽  
Dry Milk ◽  
Antibody Reaction

Abstract A new enzyme immunoassay (EIA) method for detection of motile and non-motile Salmonella was examined in a multilaboratory collaborative study. This method uses a proprietary formulation of polyclonal antibodies to Salmonella and is controlled to maintain specificity. Sensitivity is enhanced with an additional antibody reaction designed to minimize false-negative reactions attributable to steric interference that can occur during conjugate binding in immunoassay procedures. Thirty-two laboratories participated in this evaluation, which included 6 food types: nonfat dry milk, dry egg, black pepper, soy flour, chocolate, and ground poultry. Of the 1020 samples analyzed, there was a 97.2% agreement rate between the EIA method and the AOAC/Bacteriological Analytical Manual(BAM) culture method, 967.26. False-negative rates for the 2 methods were comparable for all foods and all Salmonella levels except ground poultry, where the EIA method detected significantly more confirmed positive samples than did the AOAC/BAM method. Nineteen samples were positive by EIA but negative by the culture method, and 10 samples were negative by EIA but positive by the culture method

Use of an Elevated Temperature and Novobiocin in a Modified Enzyme-linked Immunosorbant Assay for the Improved Recovery of Salmonella from Foods

1992 ◽  
Vol 55 (10) ◽  
pp. 758-762 ◽  
Author(s):  
K. F. ECKNER ◽  
D. MOVER ◽  
W. A. LEPPER ◽  
L. FANNING ◽  
M. S. CURIALE ◽  
...  
Keyword(s):  
Elevated Temperature ◽  
Incubation Temperature ◽  
False Negative ◽  
Culture Method ◽  
Microtiter Plate ◽  
Food Samples ◽  
Elisa Method ◽  
Immunosorbant Assay ◽  
Standard Culture ◽  
Enzyme Linked Immunosorbant Assay

A modified colorimetric enzyme-linked immunosorbant assay (ELISA) for Salmonella detection was compared to the standard culture method of the Bacteriological Analytical Manual/Association of Official Analytical Chemists (BAM/AOAC) using 20 artificially contaminated foods (1,200 test samples). The modifications to the current methodology consisted of an elevated incubation temperature of 42°C for the tetrathionate selective broth and M-broth postenrichments, as well as addition of 10 μg/ml novobiocin to the M-broth. The microtiter plate as not agitated during assay incubation, and centrifugation steps were eliminated from the protocol. This modified ELISA method was at least as productive as the standard AOAC culture method for the food samples tested. No false-positive reactions were encountered. The false-negative incidence was 1.5% for the immunoassay and 5.3% by the AOAC cultural method. The incidence of agreement between the methods was 96.7%.

Enzyme-Linked Immunoassay for Detection of Listeria monocytogenes in Dairy Products, Seafoods, and Meats: Collaborative Study

1994 ◽  
Vol 77 (6) ◽  
pp. 1472-1489 ◽  
Author(s):  
Michael S Curiale ◽  
Wendy Lepper ◽  
Barbara Robison
Keyword(s):  
Listeria Monocytogenes ◽  
Dairy Products ◽  
Collaborative Study ◽  
Meat Products ◽  
Culture Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Culture Methods ◽  
Inspection Service ◽  
The U.S

Abstract A collaborative study was conducted to evaluate Listeria-TekTM, an enzyme-linked immunosorbent assay (ELISA) for detection of Listeria monocytogenes and other Listeria spp. in foods. The present ELISA method was compared to the U.S. Food and Drug Administration culture method for detection of L. monocytogenes in dairy products and seafoods and to the U.S. Department of Agriculture Food Safety and Inspection Service method for detection of L. monocytogenes in meats. Replicate samples of 6 food types (frankfurters, roast beef, Brie cheese, 2% milk, raw shrimp, and crab meat) inoculated with L. monocytogenes and uninoculated control samples were analyzed by the collaborators. L. monocytogenes was identified in 593 samples by the ELISA method and in 574 samples using culture procedures. Identical results were obtained for 506 positive samples and 419 negative samples using the ELISA and culture methods for an overall agreement rate of 85.6%. The enzyme-linked immunoassay for detection of L. monocytogenes in dairy, seafood, and meat products has been adopted first action by AOAC INTERNATIONAL.

scholarly journals TECRA® Unique™ Test for Rapid Detection of Salmonella in Food: Collaborative Study

2001 ◽  
Vol 84 (2) ◽  
pp. 416-430 ◽  
Author(s):  
Denise Hughes ◽  
Angela E Dailianis ◽  
Louise Hill ◽  
Deborah A McIntyre ◽  
Aimee Anderson ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Collaborative Study ◽  
Milk Powder ◽  
Reference Method ◽  
Culture Method ◽  
The United States ◽  
Soy Flour ◽  
Milk Chocolate ◽  
Chi Square ◽  
Chi Square Test

Abstract The TECRA® Unique™ Salmonella test uses the principle of immunoenrichment to allow rapid detection of Salmonellae in food. A collaborative study was conducted to compare the TECRA Salmonella Unique test with the reference culture method given in the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Three food types (milk powder, pepper, and soy flour) were analyzed in Australia and 2 food types (milk chocolate and dried egg) were analyzed in the United States. Forty-one collaborators participated in the study. For each of the 5 foods at each of the 3 levels, a comparison showed no significant differences (p ≥ 0.05) in the proportion of positive test samples for Unique and that for the reference method using the Chi-square test for independence with continuity correction.

scholarly journals Salmonella in Selected Foods by VIDAS® Immuno-Concentration Salmonella Plus Selective Plate Method (Hektoen Enteric, Xylose Lysine Desoxycholate, Bismuth Sulfite): Collaborative Study

2002 ◽  
Vol 85 (3) ◽  
pp. 593-608 ◽  
Author(s):  
Wendy A Lepper ◽  
Ann M Schultz ◽  
Michael S Curiale ◽  
Ronald L Johnson ◽  
J Agin ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Culture Method ◽  
Black Pepper ◽  
Soy Flour ◽  
Plate Method ◽  
Milk Chocolate ◽  
Dry Milk ◽  
Selective Plate ◽  
Nonfat Dry Milk ◽  
Whole Egg

Abstract The VIDAS Immuno-concentration Salmonella (ICS) plus selective plate method (Hektoen enteric, xylose lysine desoxycholate, bismuth sulfite) method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1297 of the 1455 samples. Of the 158 samples not in agreement, 82 were VIDAS ICS plus selective plate-positive and BAM/AOAC-negative, and 76 were VIDAS ICS plus selective plate-negative and BAM/AOAC-positive.

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