Enzyme-Linked Immunosorbent Assay of Mycotoxins Using Nylon Bead and Terasaki Plate Solid Phases

1984 ◽  
Vol 47 (4) ◽  
pp. 305-308 ◽  
Author(s):  
J. J. PESTKA ◽  
F. S. CHU
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Linear Response ◽  
Screening Method ◽  
Detection Limits ◽  
Microtiter Plate ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Solid Phases ◽  
New Procedures

Nylon beads and Terasaki plates were tested as solid phases for the enzyme-linked immunosorbent assay (ELISA) of the mycotoxins aflatoxin B1 (AFB1), aflatoxin M1 (AFM1) and T-2 toxin. Both methods had detection limits comparable to that of mycotoxin microtiter plate ELISAs. Using the nylon bead ELISA, ELISA competition curves for AFB1, AFM1 and T-2 toxin exhibited linear response between 1.0 to 100, 0.1 to 100, and 0.1 to 10.0 ng/ml, respectively. Response ranges for Terasaki plate ELISAs of AFB1, AFM1 and T-2 toxin were 1.0 to 50, 0.05 to 0.50, and 0.5 to 1.0 ng/ml, respectively. The new procedures did not require specialized instrumentation and may be used as an economical screening method for mycotoxins in the field and to diagnose certain mycotoxicoses.

Simultaneous Determination of Aflatoxin B1 and Aflatoxin M1 in Food Matrices by Enzyme-Linked Immunosorbent Assay

2012 ◽  
Vol 6 (3) ◽  
pp. 767-774 ◽  
Author(s):  
Wenxiao Jiang ◽  
Zhanhui Wang ◽  
Greta Nölke ◽  
Jing Zhang ◽  
Lanlan Niu ◽  
...  
Keyword(s):  
Simultaneous Determination ◽  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Food Matrices

scholarly journals Quantitation of Aflatoxin B1-N7-Guanine Adduct in Urine by Enzyme-Linked Immunosorbent Assay Coupled with Immunoaffinity Chromatography

1997 ◽  
Vol 80 (5) ◽  
pp. 1013-1022 ◽  
Author(s):  
Tfflrumurthy Vidyasagar ◽  
Vadapalli Vyjayanthi ◽  
Nayak Sujatiia ◽  
Beedu Sashidhar Rao ◽  
Ramesh Venkataramana Bhat
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Polyclonal Antibodies ◽  
Immunoaffinity Chromatography ◽  
Competitive Elisa ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column ◽  
Indirect Competitive Elisa

Abstract IndiaA specific and sensitive method to quantitate aflatoxin B1-N7-guanine adduct in urine samples by immunoaffinity chromatography coupled with indirect competitive enzyme-linked immunosorbent assay (ELISA) is reported. A novel in vitro method to synthesize an antigen (bovine serum albumin-guanine-aflatoxin B1) and its use to produce polyclonal antibodies specific to the hapten aflatoxin B-N7-guanine are discussed. An indirect competitive ELISA developed to quantitate aflatoxin adduct showed a 50% inhibition at 15.6 pmol aflatoxin B1-N7-guanine (y=66.73+ (-19.8)x, r=-0.997). Interference by aflatoxin B1 was less than 5%, and no interference by guanine, aflatoxin Gι and aflatoxin M1 was observed. An immunoaffinity column was also developed, by using these polyclonal antibodies, for single-step purification of aflatoxin B1-N7-gua-nine. The immunoaffinity column and the indirect competitive ELISA were evaluated and validated by quantitation of aflatoxin B1-N7-guanine in urine from rats dosed with aflatoxin B1 (1 mg/kg body weight). Spiking studies with standard aflatoxin B1-N7-guanine adduct at 2 and 4 μg/mL phosphate- buffered saline gave 96 and 100% recoveries, respectively, for immunoaffinity cleanup column. The method also was tested successfully for quantitating aflatoxin B1-N7-guanine adduct in spiked human urine. Recoveries of the adduct were 79-90%.

Immunoassays for Analysis of Mycotoxins

1984 ◽  
Vol 47 (7) ◽  
pp. 562-569 ◽  
Author(s):  
FUN SUN CHU
Keyword(s):  
Ochratoxin A ◽  
Aflatoxin B1 ◽  
Recent Progress ◽  
Biological Fluids ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Protein Conjugates ◽  
Advantages And Disadvantages ◽  
The Past

During the past few years, several laboratories have prepared specific antibodies against aflatoxins B1, M1, B2a and Q1, ochratoxin A, T-2 toxin, and zearalenone. These antibodies were obtained from rabbits after immunizing with various mycotoxin-protein conjugates. With the availability of these antibodies, specific, simple and sensitive radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) procedures for monitoring mycotoxins and their metabolites in foods, feeds and body fluids have been developed. In this review, details are presented for the preparation of antibodies and the application of RIA and ELISA to determine aflatoxins B1 and M1, ochratoxin A and T-2 toxin in corn, peanuts, milk and other biological fluids. The sensitivity of ELISA for analysis of these mycotoxins in foods varied from 0.1 μg/L for aflatoxin M1 in milk to 5 μg/kg of aflatoxin B1 in peanuts. The advantages and disadvantages of ELISA for monitoring mycotoxins in foods and feeds are discussed. In addition, a description of recent progress on simplified clean-up procedures which may increase the sensitivity of immunoassays is presented.

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

Competitive Direct Enzyme-Linked Immunosorbent Assay for Detection of Sulfamethazine Residues in Swine Urine and Muscle Tissue

1988 ◽  
Vol 71 (6) ◽  
pp. 1137-1140 ◽  
Author(s):  
Deborah E Dixon-Holland ◽  
Stanley E Katz
Keyword(s):  
Horseradish Peroxidase ◽  
Muscle Tissue ◽  
Immunosorbent Assay ◽  
Microtiter Plate ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sensitive Assay ◽  
Saline Extract ◽  
Swine Urine ◽  
Phosphate Buffered Saline

Abstract A sensitive assay for the detection of sulfamethazine in swine urine and muscle tissue using a direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed. Undiluted urine or a phosphate-buffered saline extract of pork muscle tissue is mixed with an enzyme-labeled conjugate of sulfamethazine and horseradish peroxidase. The mixture is added to wells of a microtiter plate coated with antibody to sulfamethazine. After the test system is incubated, washed, and re-incubated with substrate and the reaction is stopped, the absorbance is measured at 405 nm. Levels of sulfamethazine as low as 20 ng sulfamethazine/g muscle tissue and 10 ng sulfamethazine/ mL swine urine were detected and estimated

Application of ELISA to Retail Survey of Aflatoxin B1 in Peanut Butter

1986 ◽  
Vol 49 (10) ◽  
pp. 792-795 ◽  
Author(s):  
BHANU P. RAM ◽  
L. PATRICK HART ◽  
RICHARD J. COLE ◽  
JAMES J. PESTKA
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Coefficient Of Variation ◽  
Simple Procedure ◽  
Peanut Butter ◽  
Competitive Elisa ◽  
Routine Screening ◽  
Enzyme Linked Immunosorbent Assay

A simple procedure was devised for the routine screening of aflatoxin B1 (AFB1) in peanut butter using enzyme-linked immunosorbent assay (ELISA). Peanut butter samples (5 g) were artificially contaminated with AFB1 and extracted by blending with 25 ml of 55% methanol and 10 ml of hexane. The extract was filtered and aqueous filtrate analyzed by a direct competitive ELISA. Recovery of AFB1 added to peanut butter samples ranged from 85 to 112%, with an average inter-well coefficient of variation of 18.4%. The inter-assay coefficient of variation was 22.7%. Using this procedure, only 3 of 63 commercial samples of peanut butter had detectable levels (>5.0 μg/kg) of AFB1.

Determination of Total Aflatoxin Levels in Peanut Butter by Enzyme-Linked Immunosorbent Assay: Collaborative Study

1992 ◽  
Vol 75 (4) ◽  
pp. 693-697 ◽  
Author(s):  
Alan L Patey ◽  
Matthew Sharman ◽  
John Gilbert
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Peanut Butter ◽  
Collaborative Study ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Aflatoxin ◽  
Aflatoxin Level ◽  
Relative Standard

Abstract Laboratories in Australia, Japan, Spain, the United Kingdom, and the United States participated in a collaborative study to evaluate a commercial enzyme- linked immunosorbent assay for the determination of total aflatoxin. Collaborators were sent 10 randomly numbered samples (5 blind duplicates) of roasted peanut butter. Two pairs were "blank" peanut butters to which aflatoxin B1, B2, G1, and G2 standards had been added. The other 3 pairs of peanut butters were 1 low aflatoxin level sample and 2 naturally contaminated samples. The assay is based on indirect competition. Test samples containing (free) aflatoxin, added to aflatoxin-coated microwells, compete for specific monoclonal rat anti-aflatoxin. As the concentration of aflatoxin in the test samples increases, the amount of rat antiaflatoxin binding to the aflatoxin attached to the well decreases. After a wash step to remove unbound material, the amount of rat anti-aflatoxin bound to the well is determined by its reaction with peroxidase conjugated rabbit anti-rat globulin. Bound peroxidase activity is then determined by the addition of a substrate, whose color development is inversely proportional to the aflatoxin concentration and is measured by absorbance. Coefficients of variation (CV) for total aflatoxin concentrations, for mean levels of 9,30, and 89µg/kg, were between 28 and 37% for the low level and 2 naturally contaminated samples, which contained mainly aflatoxin B1. CVs for the spiked samples were lower (24-25%) for mean levels of 11 and 20 µg/kg; recoveries were 84 and 89%, respectively. Ranges for relative standard deviations for repeatabilty and reproducibility were 9-30% and 25-37%, respectively. The method has been adopted first action by AOAC International.

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