Look at the factors that affect the detection of the kit - Database & Sql Blog Articles

Test - uppercase JPG - memory 146K

Kaixin micro test

Test probe P100-M3

EL-C1600N100013-B

The quality assurance of elisa kit is a complicated process, and many important aspects affect the detection effect:

First, the impact of methodology

The ELISA assay modes include the following: double antibody sandwich method, indirect method, double antigen sandwich method, IgM antibody capture method, competitive inhibition method, in which the competition inhibition method (HBeAb, HBcAb, etc.) is unfair due to the operation time difference. The effects of competition and other factors, the results are less repeatable, qualitative

The amount is difficult to control.

Second, the reagent factor

Different batches of ELISA reagents are difficult to ensure the same quality during the production process. Even if the results are different through batch inspection, it is necessary to select and order long batch reagents and ensure storage conditions. Strict implementation of this standard can avoid the complicated process of newly establishing a quality control system and re-evaluating the reagent due to the change of the reagent batch number, and can ensure the stability of the result; for a reagent with a short period of time and low usage rate, when a small amount of reagent is dispensed, For each use, take the dispensing part to avoid the failure of the reagent caused by repeated freezing and thawing.

1. Avoid direct contact with the stop solution and substrates A and B. Once exposed to these liquids, rinse with water as soon as possible.

2. Do not eat, drink, smoke or use cosmetics during the experiment.

3. Do not use the mouth to absorb any ingredients in the kit.

Third, the sample factor

Specimen interference factors include endogenous interfering factors and exogenous interfering factors. The former includes rheumatoid factor, complement, heterophilic antibody, autoantibody, lysozyme, etc. The latter includes specimen hemolysis, specimen contamination by bacteria, and specimen storage time. Excessive length, incomplete coagulation of specimens, repeated freezing and thawing of frozen specimens, etc. Fixing an antigen or antibody in a process is called coating. In other words, the coating is the process by which the antigen or antibody binds to the surface of the solid support. The protein and polystyrene solid phase carrier are bound by physical adsorption, relying on the interaction between the hydrophobic group on the structure of the protein molecule and the hydrophobic group on the surface of the solid support. This physical adsorption is non-specific and is affected by the molecular weight, isoelectric point, concentration, etc. of the protein. The adsorption capacity of the carrier for different proteins is different. Macromolecular proteins Smaller molecular proteins usually contain more hydrophobic groups, so they are more easily adsorbed onto the surface of the solid support. IgG has a strong adsorption force on solid phase such as polystyrene, and its linkage occurs mostly on the Fc segment, and the antibody binding site is exposed to the outside, so the coating of the antibody is generally carried out by direct adsorption. Most protein antigens can also be coated in a similar manner to antibodies. Non-protein antigens that are not readily adsorbed on a polystyrene carrier can be coated in a special manner. For example, when detecting an anti-DNA antibody, DNA is required as a coating antigen, and a general-purpose solid carrier generally cannot directly bind to a nucleic acid. The polystyrene plate can be first irradiated with ultraviolet rays (for example, a 30 W ultraviolet lamp, irradiated for 75 hours at 75 cm) to increase its adsorption performance. The solid phase carrier is pre-coated with a basic protein such as polylysine or protamine, and the binding force of the nucleic acid can also be improved. also may

The avidin biotin system is used for indirect coating, that is, the avidin is first coated with the carrier, and then the biotinylated DNA is added. The coating method is uniform and firm, and has been expanded and applied to quantitative determination of various antigenic substances. .