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Human C-reactive protein colloidal gold immunochromatographic assay quantitative test paper

A technology of reactive protein and detection test paper, which is applied in the field of quantitative detection of human C-reactive protein, can solve the problems of complex reagents, slow reporting speed, complicated operation, etc., and achieves good stability and specificity, improved detection efficiency, and improved operation process. Simple and fast effects

Inactive Publication Date: 2010-11-17
沈鹤柏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The technical problem to be solved by the present invention is to provide human C-reactive protein colloidal gold immunochromatography quantitative detection test paper, to overcome the existing human C-reactive protein colloidal gold quantitative detection technology with complicated reagents, cumbersome operation and relatively slow reporting speed Defects

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation of colloidal gold-CRP antibody complex

[0049] The mouse anti-CRP monoclonal antibody was labeled on the surface of the prepared 20nm colloidal gold particles, centrifuged at low temperature, resuspended to the original volume with borate buffer, concentrated again by centrifugation to 1 / 10 of the original volume, and stored at 4°C.

[0050]

Embodiment 2

[0052] Preparation of human C-reactive protein (CRP) immunochromatographic detection card

[0053] (1) Preparation of sample pad: Glass cellulose membrane was selected as the sample pad material, soaked in sample pad treatment solution (PBS, containing BSA, sucrose, PVP, Tween), and dried at 37°C for later use.

[0054] (2) Preparation of conjugation pad: Glass cellulose membrane was used as the conjugation pad material, and the colloidal gold-CRP antibody complex prepared above was redispersed in colloidal gold diluent (Tris buffer solution, containing sucrose, BSA, PEG20000, Tween ), and sprayed on the bonding pad, and dried at 37°C.

[0055] (3) Spotting on nitrocellulose membrane (NC membrane): Spray the corresponding antibody on different positions of the NC membrane with a membrane spotter, from the lower end to the upper end are the quality control line and the quantitative line, and dry at 37°C.

[0056] (4) Assembly: Paste the absorbent paper, NC film, binding pad, a...

Embodiment 3

[0059] Quantitative detection of human CRP serum

[0060] (1) Use normal human serum as the diluent for the CRP antigen standard product to prepare serial concentration standard products: 0mg / mL, 5mg / mL, 10mg / mL, 20mg / mL, 30mg / mL, 50mg / mL, 80mg / mL, 100mg / mL, 120mg / mL, 160mg / mL, 200mg / mL.

[0061] (2) Dilute the above-mentioned standards of each concentration according to a fixed ratio, take 0.05mL dropwise into the sample hole of the test card, and read the ratio of the signal intensity of the quantitative line to the quality control line through the quantitative detection device after 10 minutes, that is, T / C value (each sample was measured 3 times with 3 test cards, and the average value was taken), and the corresponding standard curve was drawn.

[0062] (3) Use the above standard curve to detect clinical samples, and input the a and b values ​​in the drawn standard curve y=ax+b into the quantitative detection device to directly obtain the content of CRP in the actual sa...

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Abstract

The invention relates human C-reactive protein (CRP) colloidal gold immunochromatographic assay quantitative test paper, which comprises a sample pad, a bonding pad, a nitrocellulose membrane and absorbent paper mutually lapped on a bottom plate with an adhesive in turn. A colloidal gold-labeled CRP antibody complex is coated on the bonding pad; and a detection line and a quality control line are bonded on the nitrocellulose membrane, and are antibody-enveloped lines. During detection, the samples to be detected contact the lower end of the test paper and are subjected to chromatography along the test paper; if the quality control line and a quantitative line do not develop, the test is ineffective; if only the quality control line develops, the CRP concentration is lower than the lowest detection concentration; and if both the quality control line and the quantitative line become red, the content of the CRP is read through a quantitative detection device. The test paper is characterized in that the quality control line and the quantitative line are respectively arranged from the lower end to the upper end of the test paper along the sample chromatography direction.

Description

[0001] technical field [0002] The invention belongs to the field of quantitative detection of human C-reactive protein, in particular to the quantitative detection of human C-reactive protein by colloidal gold immunochromatography. [0003] Background technique [0004] C-reactive protein (C-reactive protein, CRP) was discovered by Tillet and Francis in the serum of some acutely ill patients in 1930. It is a ring-shaped five-globular protein belonging to Oligomeric calcium-binding proteins with a relative molecular mass of about 120,000. It consists of five identical The monomers of the drug are composed of non-covalent bonds, which are synthesized by the inflammatory lymphokines interleukin-6, interleukin-1 and tumor necrosis factor stimulated by liver epithelial cells. It is named for its ability to react with the capsular C polysaccharide of Streptococcus pneumoniae. CRP is primarily used as an indicator of bacterial infection and disease activity. When there is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/52G01N21/79
Inventor 沈鹤柏邢国杰
Owner 沈鹤柏
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