Analysis of the causes of the results of enzyme-linked immunosorbent assay and solutions - Database & Sql Blog Articles

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Analysis of the causes of the results of enzyme-linked immunosorbent assay and solutions

Enzyme-linked immunosorbent assay is to find Shanghai Jinma Biotechnology Co., Ltd., our company's many years of research on ELISA test has been widely used in clinical practice with high sensitivity and good specificity, but all aspects of the operation are tested. The detection effect has a great influence. If you do not pay attention, it may lead to incomplete color, flower board and other results. The reasons and solutions for the problems that often occur in each part of the operation are summarized below, in order to bring some inspiration to the peers and improve the quality of the test.

Steps

Possible Causes

Solution

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Agent

Select a good quality test reagent, strictly follow the reagent instructions, and equilibrate the reagent at room temperature for 30-60 minutes before the operation.

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1. If the serum or plasma specimens are not well separated, the sample is loaded;
2. In manual operation, too much sample plate will cause too long waiting time before loading into the incubator (especially when the indoor temperature is high);
3. When the specimen is added and the enzyme reagent is added, the enzyme splashes out of the pore.

1. The specimen is serum: it is best to store the blood naturally for 1-2 hours, then centrifuge at 3000rmp for 15 minutes; the specimen is plasma: blood specimen collection tube containing anticoagulant must be used, and the blood collection tube must be inverted immediately after blood collection. 5-10 times, after a period of time, centrifuge at 3000 rpm for 15 minutes; if it is tested within a few days, it can be placed in a refrigerator at 2-8 °C. If it is to be stored, it should be placed in a low temperature refrigerator at -20 °C.
2. Add the sample to the incubator in time.
3. After adding the enzyme reagent, use a blotting paper to gently blot and dry on the surface of the microplate.
4. If using AT or other fully automatic loading, it is best to choose FAME or other post-processing instruments plus enzyme reagents.
5. When there are many specimens, please operate in batches.

Incubate

1. When the incubation is not attached or capped, the specimen or diluent is evaporated and adsorbed on the wall of the hole, which is difficult to clean thoroughly;
2. The incubation time is artificially extended, resulting in non-specific binding surrounding the reaction well, which is difficult to clean thoroughly.

1. affixing or capping;
2. Follow the instructions to strictly control the operating time.

Washing board

1. Wash the board by hand, and the liquid crosses between the hole and the hole.
2. When the plate is washed by the semi-automatic washing machine, the amount of washing liquid is insufficient, resulting in incomplete washing; the washing plate is clogged, the suction is not complete; the washing plate is not smooth, resulting in poor washing effect.
3. Excessive reaction plates cause long waiting time for washing.

1. Ensure that the washing liquid fills all the holes, and the washing plate needle is unblocked. After washing the plate, it is best to pat dry on the absorbent paper (select clean, no or less dusty absorbent material);
2. Reasonable arrangements, or use a few more washing machines.

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