Polybrene microwell plate pre-transfusion detection method

A detection technology and microplate technology, applied in the field of pre-transfusion detection of polybrene microplates

Inactive Publication Date: 2017-05-24

重庆市涪陵三海兰陵有限责任公司

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AI-Extracted Technical Summary

Problems solved by technology

However, the reaction time of this method is 1 hour, and at the ...

Method used

With reference to 1-4 figure, the present invention is by improving manual polybrene method, first changes used consumables and supporting facilities, uses self-designed micro-orifice plate (Fig. 1) instead, does not need test tube and centrifuge; Second, change the concentration and amount of polybrene reagent. Polybrene is a tetramine polymer that can neutralize the negative charge on red blood cells, reduce the Zeta potential, and shorten the distance between cells. At low concentrations, when the distance between red blood cells is shortened, a reversible Non-specific erythrocyte aggregation phenomenon; when the concentration of polybrene increases, erythrocytes directly form non-specific erythrocyte aggregation. When the negative charge on the surface of red blood cells is restored, due to the repulsion of negative charges between red blood cells, the phenomenon of red blood cell aggregation disappears, but the real agglutination caused by red blood cell antigen antibody does n...

Abstract

The invention relates to an improved artificial polybrene microwell plate pre-transfusion detection method. The improved artificial polybrene microwell plate pre-transfusion detection method comprises following characteristics: 1, consumable items and auxiliary facilities are changed, self-designed microwell plates (represented by formula 1) are adopted, and no test tube or centrifugal machine is used; and 2, the concentration and the amount of polybrene reagent are changed. Polybrene is a four amine polymer, is capable of neutralizing negative charges on erythrocyte, reducing Zeta electric potential, and shortening the distance between cells. When the concentration of polybrene is low, and the distance between erythrocytes is reduced, reversible nonspecific erythrocyte condensation is caused via centrifugation; when the concentration of polybrene is increased, nonspecific erythrocyte condensation is caused directly. Then the surfaces of erythrocytes are negatively charged, erythrocyte condensation disappears because of negative charge repelling among erythrocytes, and erythrocyte condensation is caused by erythrocyte antigens and antibodies, and the affinity of antigens and antibodies doesn't fade away.

Application Domain

Biological testing

Technology Topic

Erythrocyte antigenChemistry +10

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Examples

Experimental program(1)

Example Embodiment

[0020] Referring to Figures 1-4, the present invention improves the manual polybrene method, first changes the consumables and supporting facilities used, and uses self-designed microplates ( figure 1 ), no test tube and centrifuge are needed; second, change the concentration and amount of polybrene reagent. Polybrene is a kind of tetraamine polymer, which can neutralize the negative charge on red blood cells, reduce Zeta potential and shorten the distance between cells. In the case of low concentration, when the distance between red blood cells is shortened, centrifugation can form a reversible Sexual non-specific red blood cell aggregation; when the concentration of polybrene increases, red blood cells directly form non-specific red blood cell aggregation. When the negative charge on the surface of the red blood cells is restored, the red blood cell aggregation disappears due to the negative charge repulsion between the red blood cells, but the real agglutination caused by the red blood cell antigen antibody does not disappear. The pre-transfusion detection technology of polybrene microplates established in the present invention does not require centrifugation. The main operations are as follows: A. Prepare red blood cell suspension with normal saline; B. Add 2 drops of plasma and 1 drop of red blood cell suspension to the microplate; C , Add 2ml of low ionic medium (LIM), gently shake the microplate by hand, and incubate at room temperature for 1 min ( figure 2 ); D. Add 2 drops of 0.1~0.15% polybrene solution, mix and let stand for 30s, and then gently shake the microplate with your hands for 1~2 minutes, visible red blood cell aggregation ( image 3 ); E. Add the resuspension solution, gently shake the microplate for about 1 minute, and observe the results; F. Judgment of the results: if the agglutination disperses, it is caused by non-specific agglutination; if the agglutination does not disperse, it is antigen Specific response of antibody binding ( Figure 4 ).

[0021] The above are only the preferred embodiments of the present invention and do not limit the scope of this patent. Any equivalent structure or equivalent process transformation made by using the contents of this specification and drawings, or directly or indirectly applied to other related technical fields , The same reason is included in the scope of patent protection of the present invention.

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