Reagent for rapidly detecting blood group antibody titer, preparation method, test strip and application

Abstract

The invention relates to the technical field of blood group detection, in particular to a reagent for rapidly detecting blood group antibody titer, a preparation method, a test strip and application. The reagent disclosed by the invention is red cell membrane coupled fluorescent microspheres; the reagent is prepared from the following raw materials in parts by mass: 0.2 part of fluorescent microspheres, 3000 parts of EDC and 4 parts of erythrocyte membranes. The reagent prepared by compounding the proper fluorescent microspheres, the EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and the erythrocyte membrane is high in fluorescence intensity and convenient to detect, can replace standard erythrocytes to serve as a detection reagent for blood group antibody titer, can be stored at normal temperature and does not need refrigeration after being coated on test paper, and can be used for detecting the blood group antibody titer. The red blood cell content is replaced by the fluorescent microspheres, so that the hemolysis phenomenon is avoided, and the method can be used for blood group antibody detection in a complex environment.

Description

technical field

[0001] The invention relates to the technical field of blood type detection, in particular to a reagent for rapidly detecting blood type antibody titer, a preparation method, a test strip and an application. Background technique

[0002] For patients with massive hemorrhagic shock, rapid and effective blood transfusion is the key to their treatment, and general-purpose O-type whole blood has no A and B antigens on the surface of red blood cells, so there is no need for cross-matching, saving time, but because O-type plasma contains anti- -A and anti-B antibodies, the antibody titers also vary from person to person, so when the blood recipient receives type O blood containing high titer antibodies, severe acute hemolytic transfusion reactions will occur. However, type O whole blood with low titers of anti-A and anti-B antibodies can be safely transfused across blood types. Low-titer type O whole blood (LTOWB) is currently available for non-specific emergency ...

Examples

preparation example Construction

[0045] In the present invention, the preparation method of the erythrocyte membrane preferably includes the following steps:

[0046]1) After mixing more than 3 parts of human fresh whole blood (type A blood or blood type B), centrifuge at 3000rpm for 5 minutes, discard the supernatant, then wash the red blood cells 2-3 times with PBS buffer or normal saline, and collect the red blood cells;

[0047] 2) The washed erythrocytes are mixed with the lysate at a volume ratio of 1:1, left to stand for 30 minutes after mixing, centrifuged at 10,000 rpm for 5 minutes, and the supernatant is discarded to obtain lysed erythrocytes; the lysate includes: 5wt.% dodecane Sodium sulfonate solution, 10wt.% calcium chloride solution or 1wt.% triton solution;

[0048] 3) Washing with ultrapure water: mix the lysed erythrocytes and ultrapure water at a volume ratio of 1:3, after mixing, centrifuge at 10000rpm for 5min, and collect the precipitate;

[0049] 4) The collected precipitate is repeat...

Embodiment 1

[0083] A reagent for rapid detection of blood group antibody titer, the reagent is composed of the following raw materials in parts by mass: 0.2 parts of time-resolved fluorescent microspheres (purchased from Changsha Meiniu Biotechnology Co., Ltd., the article number is RF02), 3000 parts of EDC and red blood cells Film 4 parts.

[0084] The preparation method of described reagent is made up of the following steps:

[0085]1) After mixing 3 parts of fresh human whole blood (type A blood), centrifuge at 3000rpm for 5 minutes, discard the supernatant, then wash the red blood cells 2-3 times with PBS buffer, and collect the red blood cells;

[0086] 2) The washed erythrocytes were mixed with 5wt.% sodium dodecylsulfonate solution at a volume ratio of 1:1, and then allowed to stand for 30 minutes after mixing, centrifuged at 10,000 rpm for 5 minutes, and the supernatant was discarded to obtain lysed erythrocytes;

[0087] 3) Washing with ultrapure water: mix the lysed erythrocyte...

Embodiment 2

[0094] A reagent for rapid detection of blood type antibody titer similar to Example 1, the only difference is that step 1) is: after taking more than 3 parts of fresh human blood (B blood) and mixing, centrifuge at 3000rpm for 5min, and discard the supernatant , and then use physiological saline to wash the red blood cells 2 to 3 times, and collect the red blood cells.

technical field

[0001] The invention relates to the technical field of blood type detection, in particular to a reagent for rapidly detecting blood type antibody titer, a preparation method, a test strip and an application. Background technique

[0002] For patients with massive hemorrhagic shock, rapid and effective blood transfusion is the key to their treatment, and general-purpose O-type whole blood has no A and B antigens on the surface of red blood cells, so there is no need for cross-matching, saving time, but because O-type plasma contains anti- -A and anti-B antibodies, the antibody titers also vary from person to person, so when the blood recipient receives type O blood containing high titer antibodies, severe acute hemolytic transfusion reactions will occur. However, type O whole blood with low titers of anti-A and anti-B antibodies can be safely transfused across blood types. Low-titer type O whole blood (LTOWB) is currently available for non-specific emergency ...

Examples

preparation example Construction

[0045] In the present invention, the preparation method of the erythrocyte membrane preferably includes the following steps:

[0046]1) After mixing more than 3 parts of human fresh whole blood (type A blood or blood type B), centrifuge at 3000rpm for 5 minutes, discard the supernatant, then wash the red blood cells 2-3 times with PBS buffer or normal saline, and collect the red blood cells;

[0047] 2) The washed erythrocytes are mixed with the lysate at a volume ratio of 1:1, left to stand for 30 minutes after mixing, centrifuged at 10,000 rpm for 5 minutes, and the supernatant is discarded to obtain lysed erythrocytes; the lysate includes: 5wt.% dodecane Sodium sulfonate solution, 10wt.% calcium chloride solution or 1wt.% triton solution;

[0048] 3) Washing with ultrapure water: mix the lysed erythrocytes and ultrapure water at a volume ratio of 1:3, after mixing, centrifuge at 10000rpm for 5min, and collect the precipitate;

[0049] 4) The collected precipitate is repeat...

Embodiment 1

[0083] A reagent for rapid detection of blood group antibody titer, the reagent is composed of the following raw materials in parts by mass: 0.2 parts of time-resolved fluorescent microspheres (purchased from Changsha Meiniu Biotechnology Co., Ltd., the article number is RF02), 3000 parts of EDC and red blood cells Film 4 parts.

[0084] The preparation method of described reagent is made up of the following steps:

[0085]1) After mixing 3 parts of fresh human whole blood (type A blood), centrifuge at 3000rpm for 5 minutes, discard the supernatant, then wash the red blood cells 2-3 times with PBS buffer, and collect the red blood cells;

[0086] 2) The washed erythrocytes were mixed with 5wt.% sodium dodecylsulfonate solution at a volume ratio of 1:1, and then allowed to stand for 30 minutes after mixing, centrifuged at 10,000 rpm for 5 minutes, and the supernatant was discarded to obtain lysed erythrocytes;

[0087] 3) Washing with ultrapure water: mix the lysed erythrocyte...

Embodiment 2

[0094] A reagent for rapid detection of blood type antibody titer similar to Example 1, the only difference is that step 1) is: after taking more than 3 parts of fresh human blood (B blood) and mixing, centrifuge at 3000rpm for 5min, and discard the supernatant , and then use physiological saline to wash the red blood cells 2 to 3 times, and collect the red blood cells.

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