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Test strip for detecting concentration of TG (thyroglobulin) and preparation method of test strip

A technology of thyroglobulin and test strips, which is applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as no questioning, achieve strong specificity, good water absorption effect, and easy access

Inactive Publication Date: 2018-04-03
SHIKANGPEI MEDICAL TECH WUHAN CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] In the diagnosis of autoimmune diseases, many immunochromatographic test strips have appeared, but the immunochromatographic test strips based on the detection of thyroglobulin have not yet appeared in the domestic and foreign markets.

Method used

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  • Test strip for detecting concentration of TG (thyroglobulin) and preparation method of test strip
  • Test strip for detecting concentration of TG (thyroglobulin) and preparation method of test strip
  • Test strip for detecting concentration of TG (thyroglobulin) and preparation method of test strip

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Experimental program
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Effect test

preparation example Construction

[0045] The main materials used in the preparation of the above test strips

[0046] 1.1. Both TG antibody and anti-human thyroglobulin antibody were purchased from GENETEX Company in the United States; goat anti-mouse IgG was self-made; chloroauric acid: product of Sigma Company; nitrocellulose membrane: product of Sartorius Company; bovine serum albumin (BSA), poly Ethylene glycol PEG20000 is a product of Sigma Company, and other commonly used reagents are analytical reagents.

[0047] 1.2 The company obtained 300 clinical samples in relevant hospitals, including 30 samples of lymph node biopsy, serum samples, plasma samples and whole blood samples from patients diagnosed with thyroid cancer, and the TG detection value was 20-300 mg / L; 60 serum samples, 60 samples of lymphatic tissue aspiration and 60 serum samples each, with TG value <20mg / L.

[0048] 1.3 Immunochromatography result interpretation recorder: model: RH001, widely produced by Ma'anshan Ruiheng Technology Co., ...

Embodiment 1

[0050] Preparation of colloidal gold pads

[0051] The preparation of described colloidal gold pad: get 100mL colloidal gold stoste solution and use concentration to be 0.2M K 2 C0 3 Adjust the solution to pH 6.5-8.0, then add 5-20ug anti-human thyroglobulin antibody for labeling, stir at room temperature for 1 hour, block with casein or bovine serum albumin, centrifuge at 12000-13000r / min for 30 minutes, discard the supernatant Liquid, redissolved to 70mL with colloidal gold working solution, and then sprayed evenly on the polyester film with a gold spraying machine, dried the obtained polyester film at room temperature and relative humidity of 15-30%, and sealed it for future use; The colloidal gold stock solution is a colloidal gold solution with a diameter of 30 ± 5nm prepared by the chloroauric acid-trisodium citrate method; the colloidal gold working solution includes 1wt% bovine serum albumin, 0.1M Tris-HCL buffer and 20wt% Sucrose and pH 8.0, see Table 1 for details....

Embodiment 2

[0056] Preparation of nitrocellulose membrane

[0057] The TG antibody of the nitrocellulose membrane was prepared with a phosphate buffer solution with a concentration of 0.01M and a pH of 7.4 to prepare an antibody solution with a concentration of 0.5-2 mg / mL, and sprayed it with a spray amount of 1.0-2.0 uL / cm The membrane instrument draws a line on the nitrocellulose membrane as the detection line, and at the same time, prepare the antibody with a concentration of 1-2mg / mL with phosphate buffer solution containing 1% of 0.01M and pH 7.4. solution, it is used as a quality control line on the nitrocellulose membrane with a spray coating instrument at a spraying amount of 1.0-2.0uL / cm. Dry under conditioned conditions, sealed and stored for future use, see Table 2 for details.

[0058] Table 2 shows the specific values ​​of the preparation parameters of three different ratios of nitrocellulose membranes

[0059]

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Abstract

The invention discloses a test paper strip for detecting the concentration of thyroglobulin and a preparation method thereof. The test paper strip comprises strip-shaped nitrocellulose membranes arranged transversely and horizontally, and longitudinally spaced and parallel to each other on the cellulose acetate membrane. A strip-shaped detection line and a quality control line are respectively provided, wherein the end of the nitrocellulose membrane close to the quality control line is abutted against a horizontally arranged strip-shaped water-absorbing pad, and the end of the nitrocellulose membrane close to the detection line Butt against a horizontal strip-shaped colloidal gold pad, and the end of the colloidal gold pad away from the nitrocellulose membrane abuts against a horizontally horizontal strip-shaped sample pad; the colloidal gold pad contains a colloidal gold label pair anti-human thyroglobulin antibody, the detection line is coated with a detection antibody that can specifically bind to the anti-human thyroglobulin antibody, and the quality control line is coated with a detection antibody that can specifically bind to the free anti-human thyroglobulin antibody Quality control antibodies.

Description

technical field [0001] The invention relates to the field of immunoassay detection. Specifically, the present invention relates to a preparation method and application of an immunochromatographic detection test paper for detecting thyroglobulin in human body fluids, and a colloidal gold immunochromatographic method applying the principle of the double-antibody sandwich method. Background technique [0002] Thyroglobulin (TG) is a macromolecular iodinated glycoprotein synthesized by thyroid follicular epithelial cells, and is the main component of thyroid follicular colloid. Thyroid hormone (T4) synthesized by thyroid follicular epithelium and Combined with TG, it becomes conjugated thyroxine, which is stored in the follicular cavity and becomes a thyroxine pool. This form of thyroxine has no biological activity, and only under the action of proteolytic enzymes, thyroxine is released from TG. Only when it is hydrolyzed into free thyroxine (FT4) can it be biologically active....

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/532G01N33/68G01N33/574
CPCG01N33/558G01N33/532G01N33/574G01N33/6893
Inventor 朱富强
Owner SHIKANGPEI MEDICAL TECH WUHAN CO LTD
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