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Kit for detecting calcium ossein and detection method

A kit and calcineurin technology, applied in the field of immunoassay, can solve the problems of long detection time and inapplicability for clinical rapid detection, and achieve the effects of short detection time, rapid and effective removal, and efficient operation

Pending Publication Date: 2022-05-10
BEIJING HOMA BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Though this method also improves the sensitivity and the accuracy of BGP content detection to a certain extent, makes the minimum detection limit of this method be 0.5-1ng / mL, but also still exists detection time too long simultaneously (detection time is about 35-40min ) problem, so it is not suitable for the application of clinical rapid detection

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  • Kit for detecting calcium ossein and detection method
  • Kit for detecting calcium ossein and detection method
  • Kit for detecting calcium ossein and detection method

Examples

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preparation example Construction

[0047] Preparation of magnetic separation reagents

[0048] The preparation method of the magnetic separation reagent of the present application comprises the following steps: take the carboxyl magnetic beads, wash them with the first buffer for several times and then use them for later use; then add EDC reagent to activate the carboxyl magnetic beads and mix them uniformly for later use; Washing with the first buffer solution obtains the magnetic separation reagent.

[0049] Wherein, the size of the carboxyl magnetic beads may be 1-3um, such as 2um. Due to the smaller particle size of the carboxyl magnetic beads, when the carboxyl magnetic beads are placed in the container, they are in a fluid state. Therefore, in the expression of this application, the amount of the relevant solution added in the carboxyl magnetic beads is expressed as the volume of the carboxyl magnetic beads count.

[0050] The first buffer is an acidic buffer, which may be MES buffer at pH 6.0.

[0051...

preparation example

[0092] Preparation Example of Magnetic Separation Reagent

[0093] The preparation method of magnetic separation reagent specifically comprises the following steps:

[0094] Take 50uL of carboxyl magnetic beads and wash them with the first buffer solution. The first buffer solution selected in this preparation is MES buffer solution with pH 6.0. Wash the carboxyl magnetic beads 3 times, and the amount of MES buffer solution added each time is 2mL , Discard the washing solution and keep the carboxyl magnetic beads for later use. Weigh 5 mg of EDC reagent and add it to 1 mL of MES solution at pH 6.0 to obtain EDC reagent-MES solution. The content of EDC in EDC reagent-MES solution is 5 mg / mL, mix well and set aside.

[0095] All the EDC reagent-MES solution obtained above was added to the spare carboxyl magnetic beads above, so that the carboxyl magnetic beads were activated, and the final solution obtained was the magnetic separation reagent. This activation process makes the...

Embodiment

[0140] This embodiment provides a kit for detecting calcein. The kit of the present application includes R1 reagent, R2 reagent, R3 reagent, R4 reagent and calibrator. The R1 reagent is a mixture of magnetic separation reagent, anti-erythrocyte antibody at a concentration of 1.3 mg / L, tween-20 and BSA-V; the R1 reagent is contained in a reagent bottle, and the volume of the R1 reagent is 6 mL. The R2 reagent is a mixture of 0.02 mg / L horseradish peroxidase-labeled BGP monoclonal antibody, goat and mouse serum; the R2 reagent is contained in a reagent bottle, and the volume of the R2 reagent is 12 mL. The R3 reagent is a mixture of 0.02 mg / L Acridan-labeled BGP monoclonal antibody, goat and mouse serum; the R3 reagent is contained in a reagent bottle, and the volume of the R3 reagent is 12 mL. Reagent R4 is physiological saline; R4 reagent is contained in a reagent bottle with a capacity of 20mL. Calibrators include osteocalcin antigen solutions with concentrations of 0ng / mL, ...

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Abstract

The invention discloses a kit for detecting calcium ossein and a detection method, belongs to the technical field of immunodetection, and solves the problem of long time consumption of related detection methods. The kit comprises a calibrator, normal saline, a reagent R1, a reagent R2 and a reagent R3, the R1 reagent comprises a magnetic separation reagent and an anti-erythrocyte antibody; the R2 reagent comprises a BGP monoclonal antibody marked by HRP (horse radish peroxidase); the R3 reagent comprises a BGP monoclonal antibody marked by Acridan; the method comprises the following steps: S1, adding normal saline and an R1 reagent into human whole blood, reacting, and carrying out magnetic separation to obtain a reaction solution A; s2, adding a reagent R2 and a reagent R3 into the reaction solution A, and incubating to obtain a reaction solution B; s3, adding excitation liquid and spacer liquid into the reaction liquid B, and measuring; and S4, drawing a standard curve, and determining the BGP content. The kit and the method are used for detecting the BGP content in human whole blood, and have the advantages of high detection efficiency and less time consumption.

Description

technical field [0001] The invention relates to the technical field of immunoassay, more specifically, it relates to a kit and a detection method for detecting calciosin. Background technique [0002] Osteocalcin is mainly synthesized and secreted by osteoblasts and odontoblasts, and belongs to γ-carboxyglutamic acid-containing proteins. The glutamic acid group in osteocalcin can only have biological effects after γ-carboxylation, and vitamin K must participate in the carboxylation, so it is also a vitamin K-dependent protein. Osteocalcin, also known as bone gamma-carboxyglutamic-acid-containing proteins (BGP), accumulates after the peak period of bone mineralization. The BGP gene is first translated into a proosteocalcin with 88 amino acid residues. After the proosteocalcin removes the signal peptide, the γ-carboxylation recognition site binds with vitamin K-dependent carboxylase to undergo a carboxylation reaction, and then The glutamic acid group in osteocalcin is conve...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N33/58G01N33/577G01N33/537G01N21/76
CPCG01N33/68G01N33/531G01N33/581G01N33/577G01N33/537G01N21/76
Inventor 赵科
Owner BEIJING HOMA BIOLOGICAL ENG
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