The Development of a Quantitative ELISA for Aflatoxin B1 in Ground Peanuts Using Antibodies Isolated From the Yolk of a Laying Hen

Antibodies directed against Aflatoxin B1 (AFB1) were produced in laying hens, isolated from egg yolk and applied in an enzyme-linked immunosorbent assay (ELISA) for AFB1 in ground peanuts. The ELISA sensitivity was improved by reduction of the amount of the coating antigen absorbed onto the microplate well surfaces. The main aflatoxins B1, B2, G1, G2, M1, aflatoxicol, sterigmatocystin were found to cross-react in the competitive ELISA, 100, 25, 23, 4, 0, 0, 0%, respectively. Aflatoxin B1 could be reproducibly detected and quantified in spiked ground peanuts at levels greater than 5 ppb using a hexane and methanol solvent based peanut extraction protocol.

Download Full-text

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

Journal of Food Protection ◽

10.4315/0362-028x-49.10.792 ◽

1986 ◽

Vol 49 (10) ◽

pp. 792-795 ◽

Cited By ~ 13

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.

Download Full-text

Evaluation of an egg yolk enzyme-linked immunosorbent assay antibody test and its use to assess the prevalence ofMycoplasma synoviaein UK laying hens

Avian Pathology ◽

10.1080/03079450310001636318 ◽

2004 ◽

Vol 33 (1) ◽

pp. 91-95 ◽

Cited By ~ 23

Author(s):

Jennifer C. Hagan ◽

Nicholas J. Ashton ◽

Janet M. Bradbury ◽

Kenton L. Morgan

Keyword(s):

Immunosorbent Assay ◽

Laying Hens ◽

Antibody Test ◽

Egg Yolk ◽

Enzyme Linked Immunosorbent Assay

Download Full-text

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

Journal of AOAC International ◽

10.1093/jaoac/80.5.1013 ◽

1997 ◽

Vol 80 (5) ◽

pp. 1013-1022 ◽

Cited By ~ 5

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%.

Download Full-text

Evaluation of an egg yolk enzyme-linked immunosorbent assay antibody test and its use to assess the prevalence of Mycoplasma gallisepticum infection in laying hens in Italy

Italian Journal of Animal Science ◽

10.4081/ijas.2005.272 ◽

2005 ◽

Vol 4 (3) ◽

pp. 272-275 ◽

Cited By ~ 1

Author(s):

Giovanni Tosi ◽

Roberto Leonelli ◽

Marco Tamba ◽

Roberto Calabrese

Keyword(s):

Immunosorbent Assay ◽

Laying Hens ◽

Antibody Test ◽

Egg Yolk ◽

Mycoplasma Gallisepticum ◽

Enzyme Linked Immunosorbent Assay

Download Full-text

Calcium deposition in egg due to substitution of limestone by eggshell flour in feed of laying hens

Journal of the Indonesian Tropical Animal Agriculture ◽

10.14710/jitaa.43.3.257-264 ◽

2018 ◽

Vol 43 (3) ◽

pp. 257 ◽

Cited By ~ 1

Author(s):

S. Kismiati ◽

T. Yuwanta ◽

Z. Zuprizal ◽

S. Supadmo ◽

U. Atmomarsono

Keyword(s):

Laying Hens ◽

Egg Yolk ◽

Laying Hen ◽

Calcium Deposition ◽

Experimental Unit ◽

Egg Weight ◽

Tibia Bone ◽

Randomized Design

The aim of this research was to evaluate calcium deposition in egg using eggshell flour as a limestone substitute in feed. Two hundreds laying hen of Isa Brown strain of 25 weeks were used in this study. Treatments were diet with 7.5% limestone as control (T0), 2.5% limestone is substituted with eggshell flour(T1), 5% limestone is substituted with eggshell flour (T2) and limestone is substituted with eggshell flour (T3). A completly randomized design were used to allocated the treatments with 5 replications of each. Each experimental unit consists of 10 laying hens. Parameters measured were egg weight, yolk weight, albumen weight, eggshell weight, calcium of egg (yolk, albumen and eggshell), length, weight and Ca of tibia bone. The results showed that substitution of limestone with eggshell flour had significantly effect (P<0.05) on eggshell weight, Ca deposition on yolk, albumen, and Ca of tibia bone but non significantly effect on egg weight, weight and percentage of yolk, weight and percentage of albumen, percentage and Ca of eggshell, length and weight of tibia bone. In conclusion, calcium deposition in yolk was the highest in the use of 7.5% eggshell flour to substitute limestone but obtained the lowest Ca of bone , while calcium deposition in albumen was the highest in the use of 2.5% eggshell flour.

Download Full-text