Common causes and solutions in Elisa testing - Database & Sql Blog Articles

Mobile phone crystal 3.2*2.5mm 3225 26M (26.000MHZ) 7.5PF 10PPM 20PPM 30PPM

Single chip microcomputer STM32L151CCU6

Test - lowercase jpg

Kaixin micro test

There are many reagents for detecting reagents in elisa kits; there are some differences in the sensitivity and specificity of reagents produced by different manufacturers. The use of inferior reagents will inevitably lead to false positives or false negatives. Therefore, choosing high quality reagents is one of the keys to ensuring accurate results. This article will provide you with a comprehensive analysis of what problems Elisa kit will have in the experiment and provide solutions. Welcome to click to read.

Steps

Possible Causes

Solution

Selection reagent

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.

Loading

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.

Color development

1. The developer is left for a long time after preparation or use an expired color developer;
2. When the developer is added, the liquid is recirculated outside the hole.

1. The coloring agent should be prepared as far as possible before use. It should not be used without expiring the color developer. The light blue TMB color developer can be seen by the naked eye.
2. Keep the developer from flowing out while loading;
3. A, B liquid should avoid contact with metal equipment.

termination

More bubbles are generated when the stop solution is added, resulting in an increase in false positives.

Avoid air bubbles when adding stop solution.

Reading board

The bottom of the board is not clean when reading the board.

The microplate should be cleaned.

The whole process

1. Ensure that the microplate is not exposed to hypochlorous acid during the whole operation;
2. Realize the automation of ELISA test standards and effectively improve the quality of testing.

In the actual operation, in addition to selecting excellent reagents, it is necessary to strictly follow the operation steps, and at the same time, make indoor quality control and inter-room quality assessment, and test each specimen with strict working style to ensure the quality of inspection. At present, a considerable number of units in China have automatic microplate readers, which plays an important role in achieving standardized detection of ELISA and improving the quality of testing.
The company provides high-quality, affordable ELISA kits, friends in need, please call us, QQ, mobile phone to contact us! ELISA kit specifications: 96T/48T