C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent

A semi-quantitative detection and reaction protein technology, applied in the field of molecular biology, can solve the problems of inapplicability to small-scale hospitals, long detection time, etc., and achieve the effects of reducing detection application cost, sensitive response, and wide application range.

Active Publication Date: 2014-12-10
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the immunoturbidimetric method is the most common method, but the traditional CRP detection is not suitable for single-person testing, the detection time is long, and it requires a large biochemical detector, which is not suitable for small-scale hospitals

Method used

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  • C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent
  • C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent
  • C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] 1. Dilute the sample (serum or plasma) with sample diluent at 1:200;

[0054] 2. Take 40 μl of the diluted sample and add it dropwise to the sample pad on the test paper disclosed by the present invention;

[0055] 3. Stand at room temperature for 10 minutes;

[0056] 4. After 10 minutes, judge the sample test result according to the quality control line.

[0057] 1. Preparation of colloidal gold

[0058] Raw materials:

[0059] 2%HAucl 4 (1g HAucl 4 dissolved in 50ml deionized water)

[0060] 1% trisodium citrate solution (1g trisodium citrate dihydrate dissolved in 100ml deionized water)

[0061] method:

[0062] 1. Take 198ml ddH 2 O (double distilled water) in the Erlenmeyer flask, add 2ml 2% HAuCL 4 solution, mix well.

[0063] 2. Place the Erlenmeyer flask on top and heat to boil.

[0064] 3. Quickly add 5ml 1% trisodium citrate solution at one time and continue to boil.

[0065] 4. Observe the color change of the solution, from light yellow→black→purp...

Embodiment 2

[0102] The difference between embodiment 2 and embodiment 1 only lies in:

[0103] 2. Antibody labeling (gold standard solution, follow the steps below to configure step by step)

[0104] 4. Add 5 μg CRPⅡ and 10 μg rabbit IgG respectively, mix immediately after adding the antibody, and let stand for 15 minutes

[0105] 7. Finally, resuspend with 1ml gold resuspension solution pH74, 20mM Tris-Hcl (containing 0.1% casein, 5% sucrose, 1% BSA, 0.3% tween20; 0.05% NaN 3 ) to resuspend.

[0106] 3. Scribe and touch gold

[0107] dash:

[0108]1. Dilute the CRPⅠ antibody to 0.1mg / ml, and the dilution solvent is PBS (10nM-50mM, in terms of phosphate concentration, the preferred concentration here is 10mM), to obtain the reagent for marking with the CRPⅠ antibody for coating.

[0109] GAR (goat anti rabbit) antibody was diluted to 0.5mg / ml and 0.1mg / ml respectively, and the dilution solvent was PBS (10-50mM, in terms of phosphate concentration, the preferred concentration here was ...

Embodiment 3

[0114] The difference between embodiment 3 and embodiment 1 only lies in:

[0115] 2. Antibody labeling (gold standard solution, follow the steps below to configure step by step)

[0116] 4. Add 1 μg CRPⅡ and 15 μg rabbit IgG respectively, mix immediately after adding the antibody, and let stand for 15 minutes

[0117] 7. Finally, resuspend with 1ml gold resuspension solution pH10, 20mM Tris-Hcl (containing 0.5% casein, 15% sucrose, 3% BSA, 0.15% tween20; 0.05% NaN 3 ) to resuspend.

[0118] 3. Scribe and touch gold

[0119] dash:

[0120] 1. Dilute the CRPⅠ antibody to 1.5mg / ml, and the dilution solvent is PBS (10nM-50mM, based on the concentration of phosphate, the preferred concentration here is 50mM), to obtain the reagent for marking with the CRPⅠ antibody for coating.

[0121] GAR (goat anti rabbit) antibody was diluted to 0.1mg / ml and 0.5mg / ml respectively, and the dilution solvent was PBS (10-50mM, in terms of phosphate concentration, the preferred concentration he...

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Abstract

The invention discloses a C-reactive protein (CRP) semi-quantitative detection reagent and test paper using the reagent. The CRP semi-quantitative detection reagent comprises the following components: 0.1-3 mg / ml of a coating CRP I antibody mark line, 0.1-1 mg / ml of a coating goat anti rabbit antibody C1 line, 0.1-1 mg / ml of a coating goat anti rabbit antibody C2 line, a marking CRP II antibody, the content of which in per milliliter of colloidal gold is 1-10 micrograms, a marking rabbit IgG antibody, the content of which in per milliliter of colloidal gold is 5-20 micrograms, a gold re-suspension solution which is a Tris-Hcl re-suspension solution with 20mM of Tris and with the pH being 7-10, a sample gasket treatment solution which is a Tris-Hcl solution with 20mM of Tris and with the pH being 7-10, and a sample dilution solution which is a Tris-Hcl solution with 20mM of Tris and the pH being 8. The detection reagent and the test paper are convenient to use, high in detection efficiency, high in response speed, high in sensitivity and high in specificity.

Description

technical field [0001] The invention relates to a C-reactive protein detection reagent in the field of molecular biology and a test paper using the reagent, in particular to a C-reactive protein semi-quantitative detection reagent and the test paper using the reagent. [0002] The meanings of the terms involved in this patent document: [0003] 0.9% Nacl: means that every 100mL of the prepared solution contains 0.9g of Nacl. [0004] Similar formats mean similar meanings, unless otherwise stated herein. Background technique [0005] C-reactive protein, named after Tillet and Francis found in the serum of patients with acute lobar pneumonia in 1930 that it can precipitate and react with pneumococcal C polysaccharide in the presence of calcium ions, is an important acute-phase reactive protein in humans, and its concentration in the acute phase can vary. Increased by thousands of times, the half-life of CRP in the circulation is 19 hours. Human CRP is produced by the live...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/558G01N33/68
Inventor 蒋海张闻周海滨王建飞
Owner NINGBO RUI BIO TECH
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