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Colloidal gold test paper strip for detecting eosinophil derived neurotoxin

A technology of eosinophilic granulocyte and colloidal gold test paper, which is applied in the field of clinical medical detection, can solve the problems of the stage, severity, prognosis and prevention of allergic diseases, lack of effective inspection methods, delay in diagnosis and treatment of patients, and limited inspection methods. Achieve the effect of improving inspection and diagnosis efficiency, short time consumption and high sensitivity

Pending Publication Date: 2019-10-22
CHONGQING THREE GORGES MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The diagnosis of allergic diseases mainly depends on the medical history and symptoms. The clinical hospital is limited by the conditions. The examination is mainly blood test for IgE and exudate for eosinophils. However, these two tests are complicated and time-consuming, which leads to delays in the diagnosis and treatment of patients.
Moreover, due to limited inspection methods, there is no effective inspection method for the stage, severity, prognosis and prevention of allergic diseases

Method used

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  • Colloidal gold test paper strip for detecting eosinophil derived neurotoxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Preparation of coating solution containing colloidal gold-labeled mouse anti-human eosinophil-derived neurotoxin-specific antibody

[0027] 1) Preparation of colloidal gold particles with a particle size of 30 nm by trisodium citrate reduction method: take 100 mL of HAuCl aqueous solution with a mass fraction of 0.01% and heat it to boiling, then add 1 mL of trisodium citrate aqueous solution with a mass fraction of 1% while stirring , and continue to boil for 5min to obtain colloidal gold particles with a particle diameter of 30nm;

[0028] 2) Suspension preparation: add 0.6g Tris, 0.5g sodium caseinate, 0.6g PVP with a molecular weight of 360000, 10g sucrose and 0.2g PEG with a molecular weight of 20000 to distilled water, then set the volume to 200mL;

[0029] 3) Add 1 mL of the colloidal gold particles prepared in step 1) to 10 μL of 0.2 M potassium carbonate buffer, then add 30 μg of mouse anti-human eosinophil-derived neurotoxin-specific antibody, mix well and...

Embodiment 2

[0036] Test the sensitivity of the colloidal gold test strip for detecting eosinophil-derived neurotoxin in the present invention

[0037]Calf serum was used to simulate human serum, eosinophil-derived neurotoxin (EDN) was used as antigen, and the concentration of EDN was 0ng / mL, 15ng / mL, 20ng / mL, 25ng / mL, 30ng / mL, 40ng / mL , 50ng / mL, 60ng / mL, 70ng / mL and 80ng / mL of the test solution, each test solution is taken 50μL and added dropwise to 10 colloidal gold for detecting eosinophil-derived neurotoxin in Example 1 Put the sample pad on the test strip, and read the results with naked eyes after 5 minutes. The results are shown in Table 1.

[0038] EDN concentration (ng / mL) 0 15 20 25 30 40 50 60 70 80 result － － ± + + + + ++ ++ ++

[0039] Note: In the test strip, only the C line is colored, indicated by "-", and the result is negative; only one T line and C line are colored, and the color of the T line is lighter, indicated by "±", the resu...

Embodiment 3

[0042] Test the specificity of the colloidal gold test strip for detecting eosinophil-derived neurotoxin in the present invention

[0043] Calf serum was used to simulate human serum, EDN was used as antigen, and IL-2, IL-4, ECP, and MBP were used as interference antigens, and the concentrations of EDN, IL-2, IL-4, ECP, and MBP were prepared at 20ng / mL and 25ng, respectively. / mL, 30ng / mL, 40ng / mL, 50ng / mL and 60ng / mL of the test solution, 50 μL of each test solution was added dropwise to 30 eosinophil-derived neurotoxins in Example 1. Put the sample pad on the colloidal gold test strip, and read the results with naked eyes after 5 minutes. The results are shown in Table 2.

[0044] Concentration (ng / mL) 20 25 30 40 50 60 EDN ± + + + + ++ IL-2 － － － － － － IL-4 － － － － － － ECP － － － － － － MBP － － － － － －

[0045] Note: In the test strip, only the C line is colored, indicated by "-", and the result is ne...

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Abstract

The present invention relates to a colloidal gold test paper strip for detecting eosinophil derived neurotoxin and belongs to the technical field of clinical medical detection. The colloidal gold teststrip comprises a bottom plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water absorbing pad; the sample pad, the colloidal gold pad, the nitrocellulose membrane and the water absorption pad are sequentially fixed on the bottom plate in a staggered manner; the colloidal gold pad is coated with a colloidal gold-labeled mouse anti-human eosinophil derived neurotoxin specific antibody; and the nitrocellulose membrane is sequentially provided with two detection lines and one control line, wherein the two detection lines are respectively coated with mouse anti-human eosinophil derived neurotoxin specific antibodies, and the control line is coated with a rabbit anti-mouse antibody. The test paper strip has the advantages of high sensitivity and strong specificity. Thetest paper strip is used for detection, short time is consumed, detection operation is simple and convenient, special instruments and equipment and professional technical personnel are not required,and clinical examination and diagnosis efficiency can be effectively improved.

Description

technical field [0001] The invention belongs to the technical field of clinical medical detection, and in particular relates to a colloidal gold test strip for detecting eosinophil-derived neurotoxin. Background technique [0002] Eosinophils are differentiated from bone marrow hematopoietic stem cells. Once activated, eosinophils can rapidly release a large amount of pre-synthesized inflammatory mediators, thereby triggering an inflammatory response in the tissue. Current studies have determined that the specific markers of eosinophil inflammatory activation are eosinophil-derived neurotoxin (EDN) and the like. EDN has biological effects such as cell recognition, cytotoxicity, and antiviral activity. Its specific expression in allergic diseases is highly sensitive and can be used as an important monitoring indicator for diagnosis, disease activity, and prognosis. Its sensitivity is even higher than that of EDN. Monitor Eos. [0003] Immunogold labeling technique (Immunogo...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558G01N33/531
CPCG01N33/6893G01N33/6854G01N33/558G01N33/531
Inventor 邓雪松李智红刘璟游进
Owner CHONGQING THREE GORGES MEDICAL COLLEGE
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