Method and kit for cross-over lymphatic toxicity experiment

A lymphatic toxicity and experimental technology, applied in the field of cross-lymphatic toxicity experiments, can solve the problems of limited development, counting and typing errors, carcinogenicity, etc., and achieve the effects of improving sensitivity and accuracy, reducing experimental costs, and expanding the reaction system

Inactive Publication Date: 2018-10-16
SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Fluorescence staining detection method is usually through the microplate method, the material used is Terasaki microplate, which needs to be observed and judged by an inverted fluorescence microscope, and there are the following three defects: 1. Terasaki microplate is a special reaction plate, and the space of the reaction system is limited, which is not conducive to The full mixing and reaction of each experimental c

Method used

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  • Method and kit for cross-over lymphatic toxicity experiment
  • Method and kit for cross-over lymphatic toxicity experiment
  • Method and kit for cross-over lymphatic toxicity experiment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 The method and kit of cross lymphocytotoxicity experiment of the present invention

[0036] 1. Kit composition

[0037] 1. Reagents

[0038] Lymphocyte separation solution (50ml) was purchased from Tianjin Haoyang Biological Products Technology Co., Ltd.

[0039] Cell counting blank plates (20 pieces) were purchased from SHIH-YUNG Medical Equipment Company.

[0040] Rabbit complement (1 tube), One Lambda Company.

[0041] Positive serum (100 μl) was purchased from Tianjin Xiupeng Biotechnology Company.

[0042] Trypan blue staining solution or neutral red staining solution (500ul) was purchased from Beyond Biotechnology Co., Ltd.

[0043] PBS buffer (100ml)

[0044] Normal saline (50ml)

[0045] 2. Required equipment

[0046] 37°C water bath (incubator), centrifuge, ordinary optical microscope, 8-tube tube, EP tube, 1-10μl micro-sampler, etc.

[0047] 2. Detection method

[0048] 1. Specimen

[0049] Take 1 ml of venous anticoagulated whole blood ...

Embodiment 2

[0076] 1. Detection method

[0077] Except that the amount of trypan blue was increased from 2 ul to 5 ul after the reaction was completed, the rest of the steps were the same as in Example 1.

[0078] 2. Accuracy verification

[0079] 1. Take 20 cases of venous anticoagulated whole blood from transplant donors, and use the traditional fluorescent staining detection method and the method of the present invention to carry out cross-lymphocytotoxicity test detection.

[0080] 2. Test results

[0081] The test results are as follows:

[0082]

[0083] It can be seen from the above table that the detection consistency between the method of the present invention and the traditional fluorescence detection method is 100% (20 / 20), indicating that the method of the present invention can accurately detect whether there is anti-donor lymphocyte antibody in the sample to be seen.

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Abstract

The invention discloses a method for a cross-over lymphatic toxicity experiment. The method comprises the following steps: (1) taking the venous anticoagulant whole blood of a transplant donor and theserum of a transplant recipient, and separating donor lymphocytes from the venous anticoagulant whole blood of the transplant donor; (2) taking a donor lymphocyte fluid, carrying out staining and marking, and adjusting the cell concentration; (3) taking and dissolving lyophilized complement; (4) taking five reaction tubes, sequentially adding two or three of the donor lymphocytes, normal saline,positive serum and the recipient serum, carrying out uniform mixing, standing the obtained mixture, adding a fifth reagent rabbit complement, uniformly mixing the mixture and the rabbit complement, and standing the new mixture; and (5) staining the new mixture, and loading the obtained sample. The invention also discloses a detection kit for the cross-over lymphatic toxicity experiment. The methodand the kit have the advantages of low toxicity, low cost, high detection accuracy and excellent clinical application prospect.

Description

technical field [0001] The invention relates to a method and kit for cross-lymphatic toxicity experiment. Background technique [0002] The cross-lymphocytotoxicity test is the complement-dependent cytotoxicity test (CDC), which is used to determine whether the recipient has pre-existing antibodies against the donor before allogeneic organ transplantation. Pre-existing antibodies in the recipient may be produced due to multiple blood transfusions, pregnancy, secondary transplantation and other reasons. If there are cytotoxic antibodies in the recipient, it may cause hyperacute rejection after transplantation, resulting in transplant failure. Therefore, it is necessary to check whether there are anti-donor lymphocyte antibodies in the recipient's serum before transplantation to prevent hyperacute rejection. experiment. [0003] In 1969, Patel and Terasaki invented the technique of complement-dependent cytotoxicity test for the screening of acute and hyperacute rejection bef...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/569
CPCG01N33/558G01N33/56983
Inventor 杨正林刘小琦
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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