Enzyme-Linked Immunosorbent Assay of Aflatoxin Bi in Naturally Contaminated Corn and Cottonseed

1986 ◽  
Vol 69 (5) ◽  
pp. 904-907 ◽  
Author(s):  
Bhanu P Ram ◽  
L Patrick Hart ◽  
Odette L Shotwell ◽  
James J Pestka
Keyword(s):  
Correlation Coefficient ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Extraction Solvent ◽  
Sample Extract ◽  
Coefficients Of Variation ◽  
Layer Chromatography ◽  
Duplicate Sample

Abstract Naturally contaminated corn and cottonseed samples were screened for aflatoxin B1 (AFB1) by a direct competitive enzyme-linked immunosorbent assay (ELISA). Samples were blended 5 min in an extraction solvent of methanol-water-dimethylformamide (70 + 29 + 1) and filtered. Filtrates were assayed by direct competition between AFBi in the corn and cottonseed samples and AFB1-peroxidase conjugate for binding to specific antibody adsorbed to a solid phase microtiter plate. Standard curves prepared using the extract of AFB1-free corn and cottonseed samples, and extraction solvent only, showed negligible interference by the sample extract in the performance of ELISA. The AFBi content in corn and dehulled cottonseed samples as determined by ELISA ranged from 7 to 422 μg/kg and 7 to 3258 μg/kg, respectively. When ELISA estimates of AFB1 in corn were compared with values obtained by thin layer chromatography (CB method), the correlation coefficient (n = 10) was 0.95. Average interassay and subsample coefficients of variation for ELISA in corn were 21.4 and 22.0%, respectively. When ELISA estimates of AFB1 in cottonseed were compared with values obtained by liquid chromatography (Pons method), the correlation coefficient (n = 15) was 0.96. Using this ELISA, 36 duplicate sample extracts can be screened for AFB1 in less than 2 h.

Indirect Enzyme-Linked Immunosorbent Assay for Saxitoxin in Shellfish

1985 ◽  
Vol 68 (1) ◽  
pp. 13-16 ◽  
Author(s):  
Fun S Chu ◽  
Titan S L Fan
Keyword(s):  
Detection Limit ◽  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chromogenic Substrate ◽  
Rabbit Igg ◽  
Lower Detection ◽  
Peroxidase Conjugate

Abstract An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of saxitoxin (STX). Antibodies against STX were demonstrated in rabbits 5 weeks after immunizing with STX-bovine serum albumin (STX-HCHO-BSA). In the ELISA, STX-HCHO-BSA or polylysine-STX was coated onto the microtiter plate, followed by incubation with standard toxin and anti-STX antibody. The amount of antibody bound to the solid phase was determined by incubation with goat anti-rabbit IgG peroxidase conjugate and a reaction with chromogenic substrate. Competitive indirect ELISA revealed that the antiserum did not cross-react with either carbamoyl-neo-STX-suIfate or tetrodotoxin. The antibodies for STX cross-reacted with decarbamoyl- STX and neo-STX about 56% and 16% as much as they did with STX, respectively. The lower detection limits for STX, decarbamoyl-STX, and neo-STX in this sytem were about 25, 45, and 156 pg per assay, respectively. When STX added to clams or mussels was assayed, the detection limit for STX was about 50-100 ppb, and recoveries were in the range of 86.8-107%.

An Indirect Enzyme-Linked Immunosorbent Assay for T-2 Toxin in Biological Fluids

1984 ◽  
Vol 47 (12) ◽  
pp. 964-967 ◽  
Author(s):  
TITAN S. L. FAN ◽  
GUANG S. ZHANG ◽  
F. S. CHU
Keyword(s):  
Immunosorbent Assay ◽  
Biological Fluids ◽  
Microtiter Plate ◽  
Reversed Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sample Extract ◽  
Rabbit Igg ◽  
Cleanup Procedure ◽  
Significant Interference ◽  
Urine Serum

An indirect enzyme-linked immunosorbent assay (ELISA) which can detect 0.2 to 1 ng of T-2 toxin per ml in urine, serum and milk was developed. T-2 hemisuccinate was conjugated to polylysine which was then coated to a microtiter plate and incubated with rabbit anti-T-2 antibody and sample extract. The amount of anti-T-2 antibody bound to the plate was then determined by reaction with goat anti-rabbit IgG-peroxidase complex and by subsequent reaction with the substrate. Samples spiked with T-2 toxin were subjected to a simple cleanup procedure by passing them through a reversed-phase Sep-Pak catridge (C18). The recoveries of tritiated T-2 toxin added to the urine, serum and milk samples were between 71 to 90% after the cleanup step. In the ELISA, significant interference was observed when more than 5 μl of sample, without cleanup treatment, were used in each analysis. After cleanup, extracts equivalent to 50 μl of serum, urine or milk per well did not significantly interfere with the assay. The recoveries of T-2 toxin added to serum (1 to 10 ng/ml), urine (0.2 to 10 ng/ml) and milk (0.2 to 10 ng/ml) after cleanup treatment as determined by the indirect ELISA were found to be 51 to 82%, 73 to 82% and 80 to 83%, respectively.

Indirect Enzyme-Linked Immunosorbent Assay for Detection of Aflatoxin B1 in Corn and Peanut Butter

1984 ◽  
Vol 47 (4) ◽  
pp. 263-266 ◽  
Author(s):  
TITAN S.L. FAN ◽  
FUNS. CHU
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Solid Phase ◽  
Peanut Butter ◽  
Enzyme Linked Immunosorbent Assay ◽  
Corn Meal ◽  
Additional Advantage ◽  
Microtiter Plates ◽  
Sample Extract ◽  
Direct Elisa

An indirect enzyme-linked immunosorbent assay (ELISA) for aflatoxin B1 (AFB1) was developed. The method involves coating AFB1-polylysine conjugate on microtiter plates as immobilized antigen, followed by incubation with free toxin standard or sample extract and anti-aflatoxin antibody from rabbits. The amount of antibody bound to the solid phase was determined by subsequent incubation with a secondary antibody conjugated with an enzyme, i.e., goat anti-rabbit IgG-horseradish peroxidase conjugate, and reaction with the chromogenic substrate. Aflatoxins were extracted from peanut butter and corn meal samples according to the BF and CB method of the Association of Official Analytical Chemists, respectively. Extracts without column cleanup treatment were dissolved in assay buffer for subsequent ELISA. Using this technique, 79.5 to 98.6% and 68 to 97% of AFB1 added in the range of 5 to 40 (μg/kg to the corn meal and peanut butter samples were recovered, respectively. The indirect ELISA achieved the same sensitivity and specificity for AFB1 as that obtained from the direct ELISA, with an additional advantage that much less antibody (100 times less) was required for the assay.

Prostatic inhibin-like peptide quantified in urine of prostatic cancer patients by enzyme-linked immunosorbent assay.

1989 ◽  
Vol 35 (7) ◽  
pp. 1376-1379 ◽  
Author(s):  
T R Teni ◽  
A H Bandivdekar ◽  
A R Sheth ◽  
N A Sheth
Keyword(s):  
Cancer Patients ◽  
Immunosorbent Assay ◽  
Prostatic Cancer ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Recovery ◽  
The Status ◽  
Polystyrene Microtiter Plate ◽  
The Mean

Abstract This is a highly specific enzyme-linked immunosorbent assay (ELISA) for measuring prostatic inhibin-like peptide (PIP) in urine, in which we use penicillinase (EC 3.5.2.6) conjugated with PIP and, as solid phase, a polystyrene microtiter plate. We used this ELISA to measure PIP in 24-h urine specimens from men with prostatic cancer (PCa) and from age-matched controls. For prostatic cancer patients the mean +/- SEM urinary PIP of 36.1 +/- 5 micrograms/24 h was significantly (P less than 0.001) lower than the mean of 127.1 +/- 9 micrograms/24 h for the age-matched controls. PIP values for 30 samples measured by both ELISA and RIA correlated well (r = 0.985). We could detect as little as 1.56 ng of PIP in a sample. Analytical recovery of added PIP ranged from 91% to 104%. Mean CVs were 8.9% within-assay and 12.7% between-assay. We believe that this ELISA will be useful in assessing the status of PIP in men with normal and diseased prostates and in examining the function of the hypothalamus-pituitary-prostate axis.

A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

Vox Sanguinis ◽  
1983 ◽  
Vol 45 (6) ◽  
pp. 440-448 ◽  
Author(s):  
S. Spitalnik ◽  
J. Cowles ◽  
M.T. Cox ◽  
D. Baker ◽  
J. Holt ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
New Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
A New Technique

Solid-phase enzyme-linked immunosorbent assay for hepatitis E virus IgG and IgM antibodies utilizing recombinant antigens and synthetic peptides

1992 ◽  
Vol 38 (1) ◽  
pp. 175-186 ◽  
Author(s):  
George J. Dawson ◽  
Kurt H. Chau ◽  
Carlos M. Cabal ◽  
Patrice O. Yarbough ◽  
Gregory R. Reyes ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Synthetic Peptides ◽  
Hepatitis E Virus ◽  
Solid Phase ◽  
Hepatitis E ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recombinant Antigens ◽  
Igm Antibodies

Detection of specific anti-leptospiral immunoglobulins M and G in human serum by solid-phase enzyme-linked immunosorbent assay.

1980 ◽  
Vol 11 (5) ◽  
pp. 452-457 ◽  
Author(s):  
B Adler ◽  
A M Murphy ◽  
S A Locarnini ◽  
S Faine
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay
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