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Aflatoxin B1 test comparative measurements on TOMTEC and IMMULON IV.(Dynatech) ELISA plate
Ildikó Barna-Vetro
Agricultural Biotechnology Center, Gödöllõ
The binding capacity of the polystyrene plate (Tomtec) was tested and compared with Immulone polystirene plate. The testing method was the direct competitive ELISA test for aflatoxin B1 mycotoxin. In the first step the polystyrene plates were coated with antibody specific to aflatoxin B1. Aflatoxin B1 standards (500, 300,150,100, 50 ppt) and enzyme-labelled aflatoxin B1 are competing for the antibody binding sites at the same time. After an appropriate incubation period the non-reacting reagents are removed by washing. The enzyme activity of the bound enzyme-conjugate was measured by adding substrate/chromogen solution (urea peroxin/tetramethylbenzidine). The enzyme-conjugate converts the colourless chromogen into a blue coloured solution. The blue colour turns to yellow when the reaction is stopped with an acidic solution. The developed colour (absorbance at 450 nm) is inversely proportional to the aflatoxin B1 concentration in standards.
According to this test the OD values of the standards on the two plates are almost the same, that refers to the similar binding capacity of the plates.
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1 |
2 |
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4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
| A |
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AFLATOXIN B1 std. pg/ml |
Plate content: |
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| B |
|
b |
b |
b |
b |
b |
b |
b |
b |
b |
b |
b |
| C |
|
500 |
500 |
500 |
500 |
500 |
500 |
500 |
500 |
500 |
500 |
500 |
| D |
|
300 |
300 |
300 |
300 |
300 |
300 |
300 |
300 |
300 |
300 |
300 |
| E |
|
150 |
150 |
150 |
150 |
150 |
150 |
150 |
150 |
150 |
150 |
150 |
| F |
|
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
| G |
|
50 |
50 |
50 |
50 |
50 |
50 |
50 |
50 |
50 |
50 |
50 |
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Measured results on TOMTEC PS 06 01S ELISA plate |
OD 450nm |
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1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
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| A |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
| B |
0.000 |
1.452 |
1.468 |
1.457 |
1.469 |
1.471 |
1.463 |
1.458 |
1.463 |
1.459 |
1.466 |
0.000 |
| C |
0.000 |
0.132 |
0.136 |
0.132 |
0.136 |
0.129 |
0.131 |
0.130 |
0.129 |
0.131 |
0.132 |
0.000 |
| D |
0.000 |
0.230 |
0.219 |
0.221 |
0.226 |
0.223 |
0.217 |
0.229 |
0.228 |
0.219 |
0.230 |
0.000 |
| E |
0.000 |
0.426 |
0.419 |
0.420 |
0.425 |
0.439 |
0.423 |
0.431 |
0.435 |
0.428 |
0.433 |
0.000 |
| F |
0.000 |
0.701 |
0.711 |
0.706 |
0.699 |
0.716 |
0.705 |
0.701 |
0.712 |
0.709 |
0.713 |
0.000 |
| G |
0.000 |
1.196 |
1.198 |
1.201 |
1.203 |
1.214 |
1.199 |
1.194 |
1.195 |
1.197 |
1.210 |
0.000 |
| H |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
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Measured results on IMMULON IV. (Dynatech) ELISA plate |
|
OD 450nm |
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1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
| A |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
| B |
0.000 |
1.539 |
1.541 |
1.523 |
1.540 |
1.534 |
1.560 |
1.549 |
1.483 |
1.550 |
1.539 |
0.000 |
| C |
0.000 |
0.174 |
0.170 |
0.176 |
0.161 |
0.178 |
0.176 |
0.182 |
0.180 |
0.175 |
0.179 |
0.000 |
| D |
0.000 |
0.268 |
0.280 |
0.276 |
0.288 |
0.277 |
0.279 |
0.285 |
0.296 |
0.274 |
0.265 |
0.000 |
| E |
0.000 |
0.541 |
0.529 |
0.545 |
0.535 |
0.549 |
0.550 |
0.567 |
0.572 |
0.547 |
0.585 |
0.000 |
| F |
0.000 |
0.890 |
0.901 |
0.900 |
0.852 |
0.865 |
0.888 |
0.895 |
0.865 |
0.845 |
0.879 |
0.000 |
| G |
0.000 |
1.269 |
1.270 |
1.288 |
1.282 |
1.283 |
1.289 |
1.279 |
1.280 |
1.284 |
1.296 |
0.000 |
| H |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
0.000 |
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TOMTEC PS 06 01ES ELISA lemez |
IMMULON IV. (Dynatech) ELISA lemez |
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