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Erythrocyte treating, eluting and diffusing kit and preparation method thereof
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A kit and red blood cell technology, applied in the field of blood transfusion medicine, can solve the problems of complicated steps and difficult operation, and achieve the effects of less incubation time, simple operation and high efficiency.
Inactive Publication Date: 2014-03-26
JIANGYIN LIBO MEDICINE BIOTECH
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Abstract
Description
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Problems solved by technology
Many reagents need to be newly prepared before use, the steps are cumbersome, and some methods are not easy to operate successfully
Method used
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Examples
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Embodiment 1
[0022] Example 1: Preparation of direct antiglobulin test (DAT) negative red blood cells
[0023] (1) Preparation of eluent: Mix solution I and solution II according to the volume ratio of 1:1~1:8 to prepare eluent.
[0024] (2) Wash the DAT-positive red blood cells 6 times with 1:4 (volume ratio) normal saline to obtain packed red blood cells.
[0025] (3) Take one test tube, add 2ml of eluent, then add 1ml of washed DAT-positive packed red blood cells, and mix well.
[0026] (4) Incubate at room temperature for 2 minutes (not more than 2 minutes), immediately add 0.1ml solution III, mix well, centrifuge at 3000rpm × 1min, and separate packed red blood cells and supernatant.
[0027] (5) Wash packed red blood cells three times with normal saline at a volume ratio of 1:4, and prepare red blood cell suspension with a concentration of 2-4% with normal saline.
[0028] (6) Take another test tube and add 1 drop of the above-mentioned 2-4% concentration of red blood cells and 2 d...
Embodiment 2
[0029] Embodiment 2: Preparation of loose liquid
[0030] (1) Preparation of eluent: Mix solution I and solution II according to the volume ratio of 1:1~1:8, and prepare eluent.
[0031] (2) DAT-positive red blood cells were washed 6 times with 1:4 (volume ratio) normal saline to obtain packed red blood cells.
[0032] (3) Take 1 test tube, add 1~6ml of eluent, then add 1ml of washed DAT-positive packed red blood cells, and mix well.
[0033] (4) Incubate at room temperature for 2 minutes (not more than 2 minutes), immediately add 0.1ml solution III, mix well, centrifuge at 3000rpm × 1min, and separate packed red blood cells and supernatant.
[0034] (5) Transfer the supernatant to another clean test tube, add solution III dropwise, and take the color of the solution from purple to blue as the reaction end point.
[0035] (6) Centrifuge at 3000rpm × 3min, and the supernatant is the eluate that can be used for antibody detection.
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Abstract
The invention discloses an erythrocyte treating, eluting and diffusing kit and provides a method for dissociating IgG antibody sensitized on an erythrocyte membrane and application of the method. The erythrocyte treating, eluting and diffusing kit treats erythrocyte sensitized by the IgG antibody to obtain diffusion liquid containing antibody and negative erythrocyte in direct antiglobulin test (DAT), the diffusion liquid containing antibody and the negative erythrocyte in direct antiglobulin test not only can be used for classical indirect antiglobulin test and also can be used for detecting erythrocyte antigen or antibody in the diffusion liquid through a microcolumn gel card. The kit comprises the following three reagents including a solution I, a solution II and a solution III, wherein the solution I and the solution II are mixed to prepare eluant according to volume ratio of (1:1)-(1:8), and the solution III is used for judging reaction endpoint. The diffusing kit which is convenient to use and stable om quality and a preparation method of the kit are provided.
Description
technical field [0001] The present invention relates to a method for dissociating and sensitizing IgG antibodies on erythrocytes and its application. More specifically, the present invention relates to the composition, preparation method and application of a red blood cell treatment, elution and diffusion kit, and relates to the field of blood transfusion medicine. Background technique [0002] Agglutination or sensitization occurs under suitable conditions between antigens on red blood cells and antibodies in serum, and this combination is reversible. If some physical conditions (such as temperature) or chemical conditions (such as pH) are changed, the antibody will be separated from the antigen-antibody complex. This method is called a diffusion test. Dissociation test is to dissociate the antibody bound to the red blood cell membrane for the identification of ABO subtypes, the diagnosis of hemolytic disease of newborns and the identification of the antibody specificity o...
Claims
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Application Information
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