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C-reactive protein colloidal gold detection reagent card and preparation method thereof

A technology of reactive protein and detection reagents, which is applied in the field of medical detection, can solve the problems of system stability, false positives, low protein content, and difficult detection, etc., and achieve the effects of reducing the difficulty of dissolution, improving detection sensitivity, and increasing the binding rate

Active Publication Date: 2020-03-27
北京科跃中楷生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]On the one hand, colloidal gold immunochromatography has the characteristics of low detection cost and fast detection speed, but compared with detection platforms such as chemiluminescence and fluorescence radiation detection methods, The sensitivity of colloidal gold immunochromatography is low, and at the same time, due to the low content of protein (mostly composed of monomer and polymer protein) in urine, it is not easy to detect
On the other hand, after improving the detection sensitivity of colloidal gold immunochromatography for proteins, the system stability and false positive problems will follow.

Method used

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  • C-reactive protein colloidal gold detection reagent card and preparation method thereof
  • C-reactive protein colloidal gold detection reagent card and preparation method thereof
  • C-reactive protein colloidal gold detection reagent card and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 Staphylococcus aureus protein A (Protein A) affinity chromatography purifies CRP monoclonal antibody

[0042] Reagent preparation:

[0043] Equilibration / binding buffer: 0.01M pH 8.3 Tris-Nacl (0.2M NaCl);

[0044] Dissociation buffer: 0.1M pH 4.0 citric acid;

[0045] Regeneration buffer: 0.1M pH 2.7 glycine;

[0046] Dialysis buffer: 0.02M pH 7.4PBS;

[0047] pH neutralizer: 2M Tris.

[0048] Equipment preparation: Protein A column, low-temperature high-speed centrifuge, balance, peristaltic pump.

[0049] 1.1 Operation steps:

[0050] ①Sample preparation: The frozen ascites was melted at 4°C, filtered with soaked filter paper to remove oil, and then filtered through a 0.45um filter membrane to collect the supernatant;

[0051] ②Column preparation: Fill with appropriate amount of Protein A filler, rinse with pure water for 5-10 times the column volume, and rinse to remove ethanol;

[0052] ③Equilibration: Equilibrate the Protein A column with equilib...

Embodiment 2

[0061] 2.1 Preparation of scribed NC film

[0062] Scratching buffer includes the following components: 0.05M phosphate buffer (pH7.0), 95% ethanol solution and 0.1% sodium azide (Proclin-300), wherein, phosphate buffer, 95% ethanol solution, sodium azide The volume ratio is 94.9:5:0.1.

[0063] Add CRP monoclonal antibody at 350ul / mL to the centrifuge tube of the delineation buffer, mix well to obtain the CRP delineation buffer system, and coat it on the pretreated NC membrane detection line, and delineate the line section The ratio of length to liquid output is 1uL / cm;

[0064] Add the rabbit anti-mouse IgG into the centrifuge tube of the scratching buffer at 500ul / mL, mix well to obtain the rabbit anti-mouse IgG scratching buffer system, and coat it on the pretreated NC membrane quality control line, The ratio of the length of the scribing line to the liquid output is 1uL / cm.

[0065] Coat at 37°C for 2-4 hours, take it out and put it in a drying room to dry, and the dry...

Embodiment 3

[0079] Quality control line not included

[0080] 3.1 Comparison between the detection reagent card of the present invention and the prior art reagent card

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Abstract

The invention belongs to the field of medical detection, and particularly relates to a C-reactive protein colloidal gold detection reagent card as well as a preparation method and application thereof.The colloidal gold detection reagent card provided by the invention has the characteristics of low cost, no need of treatment of a detection sample, short detection time, high accuracy and the like,and can achieve the purposes of avoiding conventional blood sampling and painless detection.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a C-reactive protein colloidal gold detection reagent card and a preparation method thereof. Background technique [0002] Currently, more than 3,000 chemical substances or "metabolites" can be detected in human urine using techniques such as nuclear magnetic resonance spectroscopy, gas chromatography, mass spectrometry, and liquid chromatography. health condition. For example, when protein is detected in the urine, it means that there is more protein in the body, and the protein will pass through the kidneys and enter the urine. Therefore, through the analysis of specific substances in the urine, it can be judged whether the body is in a healthy state, and even a preliminary diagnosis of the epidemic can be made. [0003] On the one hand, colloidal gold immunochromatography has the characteristics of low detection cost and fast detection speed. However, compared wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/558
CPCG01N33/68G01N33/587G01N33/577G01N33/558G01N2333/4737
Inventor 詹先发柳静周东奇
Owner 北京科跃中楷生物技术有限公司
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