Test sample allergenicity detection method and kit thereof

A detection method and sensitization technology, which is applied in the field of medical detection, can solve problems such as the lack of technical means or technical guidance to solve the above technical problems, and achieve the effects of reducing animal pain, reducing detection costs, and reducing dosage

Active Publication Date: 2018-11-27
NANJING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] After searching, it was found that in the prior art represented by the Chinese invention patent application with the application number CN201711285287.7, the application publication number CN107966

Method used

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  • Test sample allergenicity detection method and kit thereof
  • Test sample allergenicity detection method and kit thereof
  • Test sample allergenicity detection method and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0051] Embodiment, detect the sensitization of 2,4-dinitrochlorobenzene (DNCB)

[0052] 1-Materials and reagents

[0053] 1.1 - Animal selection and preparation

[0054] Adult BALB / c mice, female, 8-12 weeks old, were provided by the Comparative Medicine Center of Yangzhou University, experimental animal production license number: SCXK (Shanghai) 2017-0007. The animals were kept in the barrier system of the Health Analysis and Testing Center of Nanjing Medical University, the license number for the use of experimental animals: SYXK (Su) 2015-0009, and humane care was given according to the 3R principles for the use of experimental animals.

[0055] 1.2- Instruments and reagents

[0056] Electronic balance (T1000, Changshu Shuangjie Testing Instrument Factory), electronic balance (AL104, Mettler), microplate reader (SpectraMax M2e, Molecular Devices), CO 2 Cell incubator (HERACELL-150, Heraeus), vernier caliper (0-150mm, Shanghai Minate Industrial Co., Ltd.), ultra-clean ben...

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Abstract

The invention relates to a test sample allergenicity detection method and a kit thereof. The detection method comprises the following steps: randomly dividing a plurality of mice into groups; measuring the thickness of ears of each mouse of different groups; entering into a first time period; within the first time period, smear an equal amount of a reagent on the ears of each mouse of different groups; entering into a second time period; when the second time period is ended, firstly measuring the thicknesses of the ears of the mice of different groups, further killing the mice, taking down lymph glands after the ears of the mice under a sterile condition, preparing a cell suspension, putting a 5-bromodeoxyuridine solution into the cell suspension, culturing, and testing the content of the5-bromodeoxyuridine by using an ELISA (Enzyme-Linked Immuno Sorbent Assay) method; and judging the allergenicity of a test substance according to limiting standards. By adopting the method, whether the test substance has positive allergenicity can be tested, intraperitoneal injection is avoided, then animals can be prevented from pain, the amount of the 5-bromodeoxyuridine can be greatly reduced,and the detection cost can be reduced.

Description

technical field [0001] The invention relates to a method for detecting the allergy of a test substance and a kit thereof, belonging to the technical field of medical detection. Background technique [0002] As far as the inventors know, when sensitizing exogenous chemicals stimulate the body, the body is prone to produce an allergic reaction, and to determine whether a chemical is sensitizing, it needs to be detected by animal experiments. [0003] The currently commonly used detection method uses the method of intraperitoneally injecting the test substance into the animal, but this will cause pain to the animal and does not conform to the "3R" principle of animal welfare. At the same time, the more expensive 5-bromodeoxyuridine (BrdU) will be used in this detection method, and the dosage of each mouse can reach 5 mg, and a detection test uses a minimum of 20 mice per group, that is, every The group requires at least 100mg of BrdU, making the entire assay cost prohibitively...

Claims

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Application Information

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IPC IPC(8): G01N33/569A61D7/00
CPCA61D7/00G01N33/56966G01N2800/7023
Inventor 环飞程洁秦珩靳苏香王玉邦张静姝严婷张成香李世芬王军董昊张璐璐胡艳辉钱雯章婉
Owner NANJING MEDICAL UNIV
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