A kind of water-based gel chromatography medium and method for detection

Abstract

The invention discloses an aqueous gel chromatography medium. The aqueous gel chromatography medium is composed of polysucrose, gelatin and water, wherein the concentration of polysucrose is 5-100g / L, and the concentration of gelatin is 1-10g / L , The aqueous gel chromatography medium can separate red blood cells and free immunoglobulins for the detection of incomplete antibodies and separate particulate antigens and antibodies for the detection of antigens and antibodies. Through the above method, the aqueous gel chromatography medium of the present invention and the method for detection can be applied to the detection of blood cells such as red blood cells, platelets, and white blood cells, and can also be used for other particulate antigens and free antigens in immunolabeling techniques. The separation of soluble proteins, such as ELISA, immunofluorescence, flow cytometry, etc., avoids the cumbersome washing process in the operation of immunological methods, greatly simplifies the operation steps, improves the detection efficiency, and can be used for the detection of a large number of samples. Water-based The raw materials of the gel chromatography medium are cheap, and the production technology requirements are low.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a water-based gel chromatography medium and a detection method. Background technique [0002] After the incomplete antibody (mainly IgG antibody) binds to the corresponding antigen of red blood cells (sensitized red blood cells), no visible agglutination reaction occurs in saline medium, but after adding anti-IgG antibody (secondary antibody), the secondary antibody Combined with the Fc segment of the incomplete antibody on the sensitized red blood cells, it can be bridged and an agglutination reaction occurs. This test is called anti-human globulin testor Coombs test. In the routine clinical antibody screening, identification and cross-matching blood, Recipients and donors with blood transfusion history, pregnancy history and transfusion of blood products should be tested for incomplete antibodies. The detection of incomplete antibodies is one of the most important contents in ...

AI Technical Summary

Problems solved by technology

In this regard, the traditional test tube anti-human globulin test was invented in 1945, and it has been used up to now and can only be used in the confirmation test of a small number of specimens. Due to its cumbersome operation, it has never become a routine clinical detection method; currently the most widely used microcolumn gel Antihuman globulin test, which does not require a washing process, but has the disadvantages of high reagent prices and false positives for fibrinogen in blood samples; in addition, various "enhanced tests", including protease, albumin, polybrene etc., because of their respective shortcomings, they have not been able to become a reliable method for detecting blood group incomplete antibodies

[0003] In 1945, scientists such as CoombsRRA believed that their blood should have anti-erythrocyte agglutinin based on the symptoms of some patients. However, after mixing these serums with red blood cells, no agglutination reaction occurred. Afterwards, agglutination occurs when it is used as a bridge. Initially, this method is only used to detect incomplete antibodies in serum. After development, it is also used to detect red blood cells that have combined antibodies and / or complements in patients, that is, the classic Coombstest. The former is indirect. Anti-human globulin test, the latter is a direct anti-human globulin test, the classic Coombstest is supposed to be a deterministic and standard test for incomplete antibody detection now and in the future, and it was also hoped to be applied to clinical routine incomplete antibodies at the beginning of its invention. Antibody detection and detection of small molecule antigens and antibodies other than red blood cells, but because anti-human spheres can combine with antibodies to sensitized red blood cells in serum and free immunoglobulin in serum, they must be washed and removed first in the conventional test tube Coombstest Free immunoglobulin that has not been combined with red blood cells, then add anti-human spheres, so as to avoid false negative reactions caused by the neutralization of anti-human spheres by non-specific immunoglobulins in the reaction solution, which requires centrifugation and washing with normal saline for at least three times, The non-specific immunoglobulin residue in the reaction solution is less than 1 / 5000 of the original amount, which leads to cumbersome and time-consuming operations, and some weak reaction antibodies cannot be detected, which cannot meet the clinical blood type compatibility test time. Therefore, this method has not been able to become a routine clinical test item

In order to solve this problem, some enhanced media such as polybrene (polybrene), polyethylene glycol (polyethyleneglycol, PEG), low ionic strength solution (lowionic strength saline, LISS) and enzyme solution have been developed. It can reduce or avoid washing procedures, reduce incubation time and enhance antigen-antibody reaction, but there are also some disadvantages: for example, polybrene method is easy to cause false negatives, and the operation process is not easy to standardize; in the test with PEG as the medium, anti-human globulin reagent The anti-complement component in the solution can cause a false positive reaction; some antibodies (anti-K) bind to red blood cells much less in a low-ionic solution than in normal saline, so testing in a low-ionic solution can lead to this Missed detection of antibodies; Enzyme treatment of red blood cells may cause false negative reactions due to the destruction of certain blood group antigens, and may also be due to the digestion of red blood cell membrane polypeptide molecules by proteases, exposing hidden antigens, and then interacting with anti-human globulin reagents False-positive reaction of heterologous antibodies or polyagglutination antibodies in serum reaction solution

Micro-column gel anti-human globulin test and solid-phase erythrocyte adsorption test can become experimental techniques for routine clinical application, but it is still impossible to completely replace the test tube method Coombstest.

At present, there is no method that can detect all blood group antibodies 100%. Some antibodies are more complex, such as enzyme-only antibodies. Therefore, when encountering complex specimens, multiple methods should be used to coordinate processing to improve the detection of incomplete antibodies. Yield rate, to ensure clinical blood safety

[0005] In the development of antibody detection technology, from agglutination test to today's various immunolabeling techniques, such as ELISA for detecting particulate antigens, immunofluorescence, flow cytometry, etc., they have never been separated from the washing process, and the reaction process involves 2-3 Each reaction procedure needs to be washed more than 3 times, the operation is cumbersome and time-consuming, and the clinical application is limited

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