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C-peptide chemiluminescence immunodetection kit, preparation method and detection method therefor

A chemiluminescent immunoassay and detection kit technology, which is applied in the field of immunoassay, can solve the problems of high cost, long reaction time, and low sensitivity, and achieve the effect of reducing reagent cost, reducing reaction time, and improving detection sensitivity

Inactive Publication Date: 2019-01-18
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problems of low sensitivity, long reaction time and high cost in existing methods for quantitative detection of C-P, the present invention provides a C-peptide chemiluminescent immunoassay kit and its preparation method and detection method

Method used

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  • C-peptide chemiluminescence immunodetection kit, preparation method and detection method therefor
  • C-peptide chemiluminescence immunodetection kit, preparation method and detection method therefor
  • C-peptide chemiluminescence immunodetection kit, preparation method and detection method therefor

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preparation example Construction

[0044] The preparation method of the C-peptide chemiluminescent immunoassay kit of the present invention mainly comprises the following steps:

[0045] Step 1. Streptavidin-coated magnetic particles

[0046] Take the magnetic particles, wash them three times with PB buffer solution with a concentration of 20mM, and dilute to 10mg / ml with PB buffer solution with a concentration of 20mM; Streptavidin was added proportionally, and after incubation at 37°C for 18-24 hours, the beads were placed on a magnetic separation rack to separate the magnetic beads; the supernatant was removed, washed three times with PB buffer with a concentration of 20 mM, and the blocking solution (20 mM PB buffer) was added. PB buffer, which contains 2-5% BSA), incubated at 37°C for 18-24h, placed on a magnetic separation rack to separate the magnetic beads; removed the supernatant, washed 3 times with PB buffer with a concentration of 20mM, and used a concentration of Dilute to 20mM PB buffer to 10mg / m...

Embodiment 1

[0063] Example 1 Preparation of C-peptide chemiluminescent immunoassay kit

[0064] 1) Streptavidin-coated magnetic particles

[0065] Take a certain amount of magnetic particles, wash them three times with PB buffer solution with a concentration of 20mM, and then dilute them to 10mg / ml with a PB buffer solution with a concentration of 20mM; Streptavidin was added proportionally, and after incubation at 37°C for 21 h, the magnetic beads were separated on a magnetic separation rack; the supernatant was removed, washed three times with PB buffer with a concentration of 20 mM, and PB buffer with a concentration of 20 mM (containing 3% BSA) as blocking solution, incubated at 37°C for 21h, placed on a magnetic separation rack to separate magnetic beads; removed the supernatant, washed 3 times with PB buffer with a concentration of 20mM, and diluted with PB buffer with a concentration of 20mM to 10 mg / ml to obtain a streptavidin-coated magnetic particle reagent, which is stored at ...

Embodiment 2

[0070] Example 2 Preparation of C-peptide chemiluminescent immunoassay kit

[0071] 1) Streptavidin-coated magnetic particles

[0072] Take a certain amount of magnetic particles, wash them three times with PB buffer with a concentration of 20mM, and then dilute them to 10mg / ml with a PB buffer with a concentration of 20mM; Streptavidin was added proportionally, and after incubation at 37°C for 18 hours, the magnetic beads were separated on a magnetic separation rack; the supernatant was removed, washed three times with PB buffer with a concentration of 20 mM, and PB buffer with a concentration of 20 mM (containing 2% BSA) as a blocking solution, incubated at 37°C for 18h, placed on a magnetic separation rack to separate the magnetic beads; removed the supernatant, washed 3 times with PB buffer with a concentration of 20mM, and diluted with PB buffer with a concentration of 20mM to 10 mg / ml to obtain a streptavidin-coated magnetic particle reagent, which is stored at 2-8°C fo...

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Abstract

The invention discloses a C-peptide chemiluminescence immunodetection kit, a preparation method and a detection method therefor, belongs to the field of the immunoassay, and solves the problems of lowsensitivity, long reaction time, and high cost of an existing C-P detection method. The kit comprises magnetic particles coated by the streptavidin; the C peptide antibody labeled by the biotin; theC peptide antibody labeled by the acridinium ester; the wash buffer; the chemoluminescence pre-trigger solution; a calibrator; and a quality control product. The detection method is as follows: the C-peptide antigen in the sample is immunologically reacted with the C-peptide antibody labeled by the biotin and the C peptide antibody labeled by the acridinium ester to form a biotin-C peptide antibody-C peptide antigen-C peptide antibody-acridinium ester solvent; the magnetic particles coated by the streptavidin are combined with the biotin to form a magnetic bead-streptavidin-biotin-C peptide antibody-C peptide antigen-C-peptide antibody-acridinium ester complex; and the concentration of the C peptide is calculated by adding the washing buffer and the like. According to the C-peptide chemiluminescence immunodetection kit, the preparation method and the detection method therefor, the sensitivity is high, the specificity is good, the reaction time is short, and the cost is low.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a C-peptide chemiluminescent immunoassay kit and a preparation method and a detection method thereof. Background technique [0002] C-peptide is composed of C-peptide B chain, C-peptide and C-peptide A chain. The C-peptide B chain is from the 1st amino acid to the 30th amino acid; the C-peptide is from the 33rd amino acid to the 63rd amino acid; the C-peptide A chain is from the 66th amino acid to the 86th amino acid. The C-peptide is connected to the B-chain and A-chain of the C-peptide through the 31st and 32nd arginines and the 64th lysine and 65th arginine. C-peptide enters the Golgi body with the microvesicles in the cytoplasm, and through the action of proteolytic enzymes, the 31st and 32nd arginine, the 64th lysine, and the 65th arginine are cut off to generate inactive C-peptide, and the A-chain and B-chain in the pro-C-peptide are connected to gener...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/543G01N33/553G01N33/532G01N21/76
CPCG01N21/76G01N33/532G01N33/54326G01N33/553G01N33/58G01N33/68
Inventor 韩美玉王立英高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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