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Near-infrared fluorescence detection kit for C-reactive protein and application of near-infrared fluorescence detection kit

A reactive protein and fluorescence detection technology, applied in the biological field, can solve problems such as the inability to detect CRP and the lack of sensitivity of detection methods, and achieve the effects of saving medical costs, low cost, and simple collection and processing

Inactive Publication Date: 2015-08-26
SHANGHAI CHEMTRON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When CRP is measured by clinical routine methods, the linear range of detection is generally 3-200mg / L. Due to the lack of sufficient sensitivity of the detection method, it is impossible to detect CRP with a lower content in the blood.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Preparation of immunofluorescent anti-C-reactive protein antibody particles:

[0050] Use 0.01mol / L, PH7.3±0.05 phosphate buffer to centrifuge and wash the fluorescent latex particles with a diameter of 150nm twice and disperse the latex particles, and add to the cleaned latex particles Anti-C Reactive Protein antibody[CRP135]; add EDCA to couple the antibody to the particles; add blocking solution to block the excess groups on the latex particles, and then add dicyclohexyl-18-crown-6. According to 100 micrograms of antibody to be labeled per milligram of microsphere latex, 100 micrograms of EDCA are used for labeling. A blocking solution equivalent to 20 micrograms of ethanolamine was added to each milligram of microsphere latex for blocking, and the final concentration of crown ether was 6 mg / ml.

[0051] 2. Preparation of the kit:

[0052] Dilute with 0.01M PBS buffer pH7.2 Anti-C Reactive Protein antibody[CRP-8](ab13426) to the concentration of 2.0mg / ml, and...

Embodiment 2

[0056] C-reactive protein near-infrared fluorescence detection experiment:

[0057] Kit detection:

[0058] The concentration of C-reactive protein standard and PBS buffer solution is 0.5pg / ml, 0.8pg / ml, 1.0pg / ml, 2pg / ml, 5pg / ml, 10pg / ml, 20pg / ml, 50pg / ml, 100pg respectively / ml, 200pg / ml, 300pg / ml, 400pg / ml, 500pg / ml, 1000pg / ml C-reactive protein standard solution, with 0pg / ml as the blank control, the blank control and each concentration of the standard were measured repeatedly for 5 Second-rate. Both the standard and the kit were equilibrated to room temperature to start detection, and 3 drops of sample (about 120-150ul) were added dropwise into the sample hole of each kit with a dropper. At 15 minutes, the fluorescence signal was detected with a near-infrared fluorescence detector, and judged with a fluorescence instrument. The detection range of the analyzer for the fluorescence signal is AD value 0-10000. A standard curve was established with the concentration of C-re...

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Abstract

The invention discloses a near-infrared fluorescence detection kit for C-reactive protein and application of the near-infrared fluorescence detection kit. The near-infrared fluorescence detection kit comprises a reagent strip positioned on a back plate, wherein the reagent strip is a sample pad, a glass cellulose membrane marked with an immune fluorescent probe, a nitrocellulose membrane coated with a C-reactive protein antibody and water absorbing paper from the sampling end in sequence. The near-infrared fluorescence detection kit disclosed by the invention can quickly and accurately detect the C-reactive protein in human whole blood, serum or plasma in combination with a near-infrared fluorescence detection technology.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a C-reactive protein (CRP) near-infrared fluorescence detection kit and its application. Background technique [0002] C-reactive protein (CRP) is synthesized by the liver with a relative molecular mass of 1.15×10 3 The pentacyclic structure of C, with calcophilic binding sites on each side, and the half-life of its molecular metabolic cycle is only 5-7 hours, therefore plays an important biological role in the immune regulation of the complement system. CRP is recognized as the most valuable typical acute phase protein. The normal plasma level is less than 5mg / L. After the body is stimulated by infection, trauma, tumor and physical and chemical factors, it rises rapidly within 5-8 hours, and the peak value can be normal within 24 hours. hundreds of times. If the infection is under control, CRP can drop rapidly within 24-48 hours and return to normal within 1 week. CRP can combine...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6809
Inventor 郭焕杰李林
Owner SHANGHAI CHEMTRON BIOTECH
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