Reasons and solutions for common problems of Weidweikang animal disease kit

The above problems arise

solution

Test sample

1 serum hemolysis, there will be false positives;

2 serum repeated freezing and thawing to reduce antibody titer, there will be false negatives;

3 serum is contaminated by bacteria after repeated freezing and thawing, or not mixed before use, resulting in poor repeatability of the experiment.

1 When collecting serum, it should be aseptically operated, requiring serum to be clear without hemolysis, and avoid using serum with severe hemolysis;

2 serum preservation time can not be too long, the serum should be fully mixed after thawing, the serum should be mixed evenly before the sample is added, and the serum after thawing should be tested once. If multiple tests are needed, it is recommended to pack a small amount of frozen. save;

3 If the serum appears turbid or precipitated, it is recommended to take the supernatant after centrifugation.

            Reagents and components in the kit

1 When the reagent expires or the reagent components of different batches of reagent kits are mixed, the absorbance value may be deviated;

2 The reagent is contaminated during use, the reagent is repeatedly warmed up, and the concentrated washing liquid is used without crystallization;

3 The use of the enzyme label incorrectly or has deteriorated will result in the experimental results not being established (no color development after the light reaction).

1 Do not use expired reagents and mix different batches of reagents to avoid multiple use of the kit;

2 Confirm whether the enzyme label is derived from the kit, and avoid the reagent to be taken back into the original bottle; do not remove the enzyme label too early;

3 Make sure to use the washing liquid that is supplied with the kit. The concentrated washing solution should be diluted with distilled water or deionized water. If there is crystal, it should be heated and dissolved before use.

             Laboratory ambient temperature control

1 laboratory temperature is too high or too low will lead to deviation of absorbance;

2 The kit was warmed up and the incubation was not carried out according to the specified temperature and time.

1 keep the laboratory temperature at 20 -25 °C , To avoid the direct light or near vent operation; the kit removed from the refrigerator, to be placed over 40min at room temperature, then returned to room temperature kits used immediately put to use after 2 -8 °C save;

2 The temperature and time of incubation and incubation should be accurate. When the incubation, the microplates should not be stacked to ensure that the temperature of each plate can be balanced.

          Labman's operational problems

1 When using the sampler, the operation is not correct or the sampler tip is in contact with the plate hole, causing pollution;

2 The washing liquid is not prepared correctly or is prepared with unqualified water, or the wrong washing liquid is used to wash the board, the number and method are incorrect; when the board is washed manually, the paper is coated with paper dust; the absorbent paper is reused, resulting in The contamination of the ELISA plate will affect the accuracy of the test results;

3 If the reagents are added in the wrong order or the experimental steps are omitted, the enzyme label will not be colored;

The loading time difference caused by the long 4 o'clock time will also affect the result.

1 In strict accordance with the instructions provided by the kit, the order and steps of reagent addition should be correct;

2 When using the sampler for sample loading, avoid the contact of the applicator tip with the plate hole. Replace the tip when sampling, and avoid adding the sample to the hole wall or generating air bubbles. When testing a large number of samples, use it. The multi-channel sampler operates in batches, and only one plate is operated at a time; firstly, all the serum to be tested is diluted, and then the plate is clicked to reduce the error caused by the delay;

3 Wash according to the specified number of washings. The washing liquid added to each hole is not less than the specified amount of liquid. Manually wash the plate without using the absorbent paper with paper dust, and avoid repeated use of the absorbent paper.

               Instrumentation problem

1 The reader of the microplate reader has an error;

2 The long-term use of the sampler is inaccurate due to mechanical wear;

3 The problem of contamination of the washing machine, blockage of the liquid head, etc. will lead to errors.

Instruments and equipment commonly used in the test, such as microplate reader, sampler, plate washer, incubator and refrigerator containing reagents, should be regularly maintained and calibrated to ensure the normal use of the equipment and avoid test errors.