ELISA is a highly sensitive test technique that combines the specific reaction of an antigen with the efficient catalysis of an enzyme on a substrate based on an immunological reaction.
1. The principle of operation of food safety testing ELISA kit:
ELISA procedure: sample collection and preservation, reagent preparation, incubation, washing, color development, colorimetry, coating, loading, enzyme-labeled antibody, termination reaction, result determination,
2. Influencing factors:
Molecular weight, stability, purification, coating concentration, antibody purity and product quality
Although the procedure of the ELISA assay is very simple, there are many factors that may affect the results of the assay, which are distributed in the various steps of the assay, especially for loading, incubating and washing. In order to help you analyze the possible causes of problems in the search, the common problems and causes are summarized in the following table.
Possible problems and possible causes in clinical ELISA assays
Possible cause of the problem (not the cause of the kit itself)
1. The weak positive control sample does not detect the time or temperature of the incubation; the color reaction time is too short; the distilled water used to prepare the buffer has problems.
2. Poor repeatability of the measurement (consistent results of the same sample twice) This is a typical problem caused by the measurement operation, including
(1) The amount of samples and reagents is inaccurate; the holes are inconsistent;
(2) The sample is added too fast, and contamination occurs between the holes;
(3) adding the wrong sample;
(4) When adding samples and reagents, add to the non-coated area in the upper part of the hole wall;
(5) Mixing components in different batches of kits;
(6) Incubation time, washing plate, and color development time are inconsistent;
(7) Contaminated debris in the hole;
(8) The microplate reader filter is incorrect;
(9) The serum sample is not completely coagulated, and the fibrin coagulation or residual blood cells appear in the reaction well, which is prone to false positive reaction.
3. Whiteboard
(The positive control does not develop color) (1) Leakage plus enzyme conjugate;
(2) There are problems in the preparation of the washing solution, such as the measuring cylinder is not clean, containing enzyme inhibitors (such as sodium azide).
(3) Leaking the developer A or B;
(4) Terminator is used as a color developer.
4. All the holes of the plate have color development (1) the plate is not clean;
(2) deterioration of the color developing liquid;
(3) the absorption of the substrate is contaminated by the enzyme;
(4) The washing solution is contaminated by enzymes and the like.
3. Key parameters and function range, corresponding price relationship
For the food detection ELISA kit, the imported kits account for a larger share of the domestic kits than the domestic kits. The domestic food detection ELISA kits are mainly on natural toxins, and the domestic food detection ELISA kits are In the detection of a variety of natural toxins. The main products on the market for early food testing ELISA are: R-biopharm, Neogen domestically produced kits are mainly: The companies that produce such food detection ELISA kits in the market in recent years have grown rapidly and account for the market share in China. A few large companies: Changsha Andi, Shenzhen Green Poetry, Sanfangyuan, Antu, Kehua, Soo, etc. These companies are more food testing ELISA kit products, but there are many previous animal enzymes free Companies that test kits are also entering the market for food safety testing.
(5) Do you need complex technical support such as on-site guidance from manufacturers?
The general food testing ELISA kit is relatively simple to operate, does not require the manufacturer to go to the door to guide, and the success rate of the experiment is also large.
(6) Whether the product is mature and the stability of the reagent, the technical support of the supplier and the quality of service such as replacement
The food detection ELISA kit reagent has matured and has good stability, and its application in the food field is still expanding. Generally, reagents that have problems in the kit, as long as the experimental report and the residual reagent are provided, the manufacturer's identification is indeed a problem of the reagent itself. , can provide return service.
(7) Transportation conditions and storage conditions, the expiration date of the product.
The enzyme-free kit should be transported at low temperature (2-8 degrees Celsius). The long-term storage should be stored at -20 degrees Celsius (preventing the effects of repeated freeze-thaw cycles on enzyme activity), and the long-term preservation should be done at 2-8 degrees Celsius.
Influencing factors, technical support
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