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Near infrared fluorescence detection kit for glycogen phosphorylase isoenzyme BB

A technology of phosphorylase isozymes and detection kits, which is applied in the direction of fluorescence/phosphorescence, measuring devices, instruments, etc., can solve the problems of rapid diagnosis and examination of unwell AMI, time-consuming detection methods, etc., and save social medical costs , low cost, easy to collect and process effects

Active Publication Date: 2015-09-09
SHANGHAI CHEMTRON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Commonly used enzyme-linked immunosorbent assay (ELIsA) and other detection methods are time-consuming and require special equipment, and are not suitable for the needs of emergency rooms and on-site rapid diagnosis of AMI.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Preparation of immunofluorescent anti-glycogen phosphorylase isozyme BB antibody particles:

[0046] 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-GPBB antibody [17B6]; 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.

[0047] 2. Preparation of the kit:

[0048] Dilute with 0.01M PBS buffer pH7.2 Anti-GPBB antibody [17B6] to a concentration of 2.0mg / ml, and crown ether was added, t...

Embodiment 2

[0052] Kit detection:

[0053] Glycogen phosphorylase isoenzyme BB synthesized by genetic engineering and PBS buffer are respectively prepared at concentrations of 0.4pg / ml, 0.6pg / ml, 0.8pg / ml, 1pg / ml, 2pg / ml, 4pg / ml, 10pg / ml, 20pg / ml, 50pg / ml, 100pg / ml, 200pg / ml, 300pg / ml, 400pg / ml, 500pg / ml, 1000pg / ml glycogen phosphorylase isoenzyme BB standard, expressed as 0pg / ml ml was used as a blank control, and the blank control and each concentration of the standard were repeatedly measured 6 times. After the standard and the kit are equilibrated to room temperature, start the detection. Use a dropper to add 3 drops of sample (about 120-150ul) into the sample hole of each kit. 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. The concentration of glycogen phosphorylase isoenzyme BB is taken as the abscissa, an...

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PUM

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Abstract

The invention discloses a near infrared fluorescence detection kit for glycogen phosphorylase isoenzyme BB and application of the near infrared fluorescence detection kit. The near infrared fluorescence detection kit comprises a reagent strip on a backboard, wherein the reagent strip comprises a sample pad, a glass fiber film marked by an immunofluorescence probe, a nitrocellulose membrane coated by the anti-glycogen phosphorylase isoenzyme BB antibody, and absorbent paper. According to the invention, by combining with the near infrared fluorescence detection technology, the detection kit can rapidly and accurately detect the glycogen phosphorylase isoenzyme BB in human whole blood, serum and plasma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a near-infrared fluorescence detection kit for glycogen phosphorylase isoenzyme BB and its application. Background technique [0002] Glycogen phosphorylase isoenzyme BB (GPBB) is an enzyme with high content in cardiomyocytes. Glycogen phosphorylase is the key enzyme of glycogen decomposition. It forms a glycogen decomposition complex in the sarcoplasmic reticulum of cardiomyocytes. This complex is particularly sensitive to glycogen decomposition caused by ischemia, which changes its composition and forms town-soluble sugar The original phosphorylase isoenzyme BB (GPBB) enters the blood through the cell membrane. [0003] Glycogen phosphorylase isoenzyme BB (GPBB) is sensitive to myocardial ischemia and hypoxia, and can be released into the blood in the early stage of myocardial injury (less than 4h), which is a good indicator for early diagnosis of acute myocardial injury. The ear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/573G01N33/533
Inventor 李林
Owner SHANGHAI CHEMTRON BIOTECH
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