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Platelet antigen antibody detecting or cross matching solid phase anti-human globulin kit

A platelet antigen and anti-human globulin technology, applied in the biological field, can solve the problems of difficulty in determining responsibility, short time for observing results, long time-consuming, etc., and achieve the effects of simple and easy method, easy automation, and easy results.

Inactive Publication Date: 2009-03-11
长春博德生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 1. Repeated washing and multiple incubation processes are required, which is cumbersome and time-consuming
[0007] 2. Only a small number of samples can be tested each time, and it is difficult to carry out the detection of a large number of samples
[0008] 3. The technical requirements are high, generally operated or demonstrated by senior technicians in the laboratory, and it is difficult to popularize the technology
[0009] 4. The observation time is short, and the experimental results cannot be saved. Once a medical dispute occurs, it is difficult to determine the responsibility

Method used

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  • Platelet antigen antibody detecting or cross matching solid phase anti-human globulin kit

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preparation example Construction

[0030] Preparation of indicator cells:

[0031] (1) Fresh O-type anticoagulated whole blood (D+) was taken and washed 3 times with normal saline.

[0032] (2) Mix equal volumes of washed packed red blood cells and human IgG (anti-D) with a titer of 16-64, and incubate in a water bath at 37°C for 30 minutes.

[0033] (3) After the above-mentioned sensitized cells were washed three times with physiological saline, they were prepared to a concentration of 0.2-0.5% with red blood cell protection solution.

[0034] d. Anti-human IgG: immunize laying hens with human gamma globulin, and extract anti-human IgG from eggs;

[0035] The preparation and extraction method of anti-human IgG: immunize laying hens with human gamma globulin, collect the eggs after the titer is qualified, and extract anti-human IgG from the eggs by repeated freezing and thawing.

[0036] e. Low ion medium: can enhance the antigen-antibody reaction;

[0037] Preparation of low-ion medium: Weigh 0.875g of NaCl...

Embodiment 1

[0047] Platelet antigen test:

[0048] The premise is that the user prepares antiserum containing platelet-specific antibodies.

[0049] 1. Equilibrate the kit to room temperature (18-25°C). Dilute 30× concentrated washing solution with distilled water at 1:29 to make working washing solution.

[0050] 2. Take out the reaction strips according to the experimental amount, mark the positive control, negative control and sample wells to be tested, and store the unused strips in a ziplock bag, add a desiccant to seal, and store at 2-8°C.

[0051] 3. Add 1 drop (50ul) of platelet suspension to be tested into the reaction well, and shake the reaction plate gently (about 10 seconds) to fully dissolve the protective agent in the reaction well.

[0052] 4. Use a plate centrifuge to centrifuge the reaction plate at 50g for 5 minutes to fix the platelets at the bottom of the microwells.

[0053] 5. Gently pour out unbound platelets, and use a dropper to add working washing solution to...

Embodiment 2

[0061] Platelet antibody test:

[0062] 1. Equilibrate the kit to room temperature (18-25°C). Dilute 30× concentrated washing solution with distilled water at 1:29 to make working washing solution.

[0063] 2. Take out the reaction strips according to the experimental amount, mark the positive control, negative control and sample wells to be tested, and store the unused strips in a ziplock bag, add a desiccant to seal, and store at 2-8°C.

[0064] 3. Add 1 drop (50 ul) of mixed type O platelet suspension for three persons to the reaction well, shake the reaction plate gently (about 10 seconds) to fully dissolve the protective agent in the reaction well.

[0065] 4. Use a plate centrifuge to centrifuge the reaction plate at 50g for 5 minutes to fix the platelets at the bottom of the microwells.

[0066] 5. Gently pour out unbound platelets, and use a dropper to add working washing solution to wash 3 times. During the washing process, gently shake the microplate, and then gent...

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Abstract

The invention discloses a blood platelet antigen antibody detection and cross-matching solid phase antihuman globulin kit, which pertains to the field of biotechnology and aims at providing a blood platelet antigen antibody detection and cross-matching solid phase antihuman globulin kit. The kit comprises a u-shaped reaction plate, blood platelets, indicator cells, antihuman IgG, low-ion medium, antiplatelet serum, negative control serum and positive control serum. The invention has high sensitivity, less time consuming (less than 1 hour) of the detection process, simple, convenient and feasible method and easily determined result, and can synchronously carry out detection on a great amount of samples and easily realize automation.

Description

Technical field: [0001] The invention belongs to the field of biotechnology. Background technique: [0002] Platelet antibodies are produced by allogeneic or autologous platelet antigens acting on the body, including alloantibodies, autoantibodies, alloantibodies, drug-dependent antibodies, and nonspecific antibodies. These antibodies act on human platelets and can lead to platelet destruction And cause fetal maternal alloimmune thrombocytopenia (FMAIT), platelet refractory transfusion (PTR), posttransfusion purpura (PTP) and other platelet immune diseases. [0003] Compared with the serological method for detecting erythrocyte antibody, the experimental method for clinical detection of platelet antibody appeared later. In 1987, Kiefel invented the monoclonal antibody-immobilized platelet antigen assay (MAIPA), which has become the "gold standard" for detecting platelet antibodies due to its high sensitivity and specificity. However, the test takes a long time; the technic...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/80
Inventor 李勇段生宝李东童军邰慧波
Owner 长春博德生物技术有限责任公司
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