Chemiluminescence detection kit for N-MID osteocalcin and preparation method of chemiluminescence detection kit

Abstract

The invention discloses a chemiluminescent detection kit for N-middle osteocalcin and a preparation method thereof. The kit includes: magnetic particles coupled with N-terminal osteocalcin capture antibody, acridinium ester-labeled N-terminal osteocalcin detection antibody, N-terminal osteocalcin series standards, chemiluminescent pre- Excitation solution A, chemiluminescence excitation solution B, cleaning solution. The kit of the invention uses the magnetic separation chemiluminescence technology as the detection means, and simultaneously combines the acridinium ester labeling technology. Compared with the existing technology for detecting N-terminal osteocalcin, this kit has the advantages of high sensitivity, high signal-to-noise ratio and rapid detection.

Description

technical field [0001] The invention belongs to the technical field of immunoassay and analysis, in particular to an N-middle end osteocalcin chemiluminescent immunoassay kit and a preparation method thereof. Background technique [0002] Osteocalcin is the most important specific non-collagen protein in the bone matrix, synthesized and secreted by osteoblasts in bones and teeth, and its binding to bone calcium depends on vitamin K. Osteocalcin molecule contains 49 amino acids with a molecular weight of about 5800D. In bone synthesis, osteoblasts produce osteocalcin, which depends on vitamin K, and vitamin D3 can promote the production of osteocalcin. The use of vitamin K antagonists can reduce the content of this protein in bone, but it does not affect its proline content, nor does it affect the mechanical strength of bone. After the production of osteocalcin, part of it enters the bone matrix, and the other part is released into the peripheral blood. Therefore, the cont...

AI Technical Summary

Problems solved by technology

Among them, although radioimmunoassay and radioimmunoassay have the advantages of high sensitivity and high specificity, there are many operating steps, special testing equipment is required, reagents are expensive, matching instruments are required and there is radioactive contamination, which makes the operation difficult. People are exposed to radiation hazards; in addition, radioactive isotopes are easy to decay, have a short validity period, and are not easy to store

The ELISA method uses horseradish peroxidase or alkaline phosphatase markers. The enzyme markers are easily inactivated, and the chromogenic substrate is easily decomposed when exposed to light. The sensitivity is low, the operation is complicated, and the repeatability is poor. It is not suitable for emergency and clinical use. Patients need to be diagnosed in time, and they have certain cross-reactions to compounds with similar structures, resulting in inaccurate test results

Although the latex-enhanced immunoturbidimetric method has the advantages of simple operation and rapidity, it has low sensitivity and poor reproducibility at low values.

[0005] CN 106353106 A (2017.01) discloses a kit for detecting osteocalcin by chemiluminescence, which uses alkaline phosphatase to label osteocalcin monoclonal antibody. The disadvantage of using alkaline phosphatase is that the enzymatic process is greatly affected by the environment : For example, disinfectant, pH, and ions will affect it and affect the measurement results, and the time for the substrate to reach the plateau is long, and the cost of the substrate is high, resulting in high detection costs