Kit for detecting osteocalcin, and using method thereof

A technology of osteocalcin and kits, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of high price, unfavorable popularization, large economic burden of patients, etc., achieve good stability, improve popularization, use, and variation small effect

Inactive Publication Date: 2018-12-21
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In China, the detection of osteocalcin is mainly based on the application of foreign Roche reagents. The price of imported reage

Method used

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  • Kit for detecting osteocalcin, and using method thereof
  • Kit for detecting osteocalcin, and using method thereof
  • Kit for detecting osteocalcin, and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A test kit for detecting osteocalcin, characterized in that it includes a calibration product, a quality control product, a solution of a biotin-labeled osteocalcin antibody, a solution of a horseradish peroxidase-labeled osteocalcin antibody, streptavidin Avidin-labeled nano magnetic particle suspension, substrate solution and washing solution;

[0034] The calibrators include osteocalcin antigen solutions with concentrations of 0 ng / mL, 10 ng / mL, 25 ng / mL, 50 ng / mL, 75 ng / mL, and 250 ng / mL, and the osteocalcin antigen was purchased from sigma, the product number is O5761; the calibrator is prepared from osteocalcin antigen and 0.1M Tris-HCl buffer with a pH value of 7.2;

[0035] The quality control products include osteocalcin antigen solutions with a concentration of 25 ng / mL and 75 ng / mL, and the osteocalcin antigen is purchased from sigma, and the article number is O5761; the quality control products are composed of osteocalcin antigen and 0.1M, Tris-HCl buffer s...

Embodiment 2

[0047] Prepare different concentrations of biotin-labeled antibody and horseradish peroxidase-labeled antibody, group the biotin-labeled antibody and horseradish peroxidase-labeled antibody according to the concentration ratio, and measure the osteocalcin concentration as follows: The luminescence values ​​at 0, 25, and 250ng / ml are shown in Table 1.

[0048] Table 1: Effect of different concentrations of anti-reagent concentration on luminescence value

[0049]

[0050] Result evaluation: When the concentrations of biotin-labeled antibody and horseradish peroxidase-labeled antibody are 1.0 μg / ml and 1.0 μg / ml respectively, the ratio of the luminescence value of the high-concentration and low-concentration calibrator is the largest, so this concentration ratio is selected as Working concentration for subsequent experiments.

[0051] According to the screened optimal concentration of anti-reagent, the effects of different sample volumes and different incubation times on the...

Embodiment 3

[0058] The method for detecting osteocalcin using the kit of the above-mentioned embodiment 1 comprises the following steps:

[0059] (1) Add 40 μL of the serum to be tested or calibrator, 60 μL of biotin-labeled osteocalcin antibody, and 50 μL of streptavidin-labeled magnetic nanoparticles to the test tube, mix well, and incubate at 37°C for 15 Minutes, the osteocalcin in the sample is captured and fixed on the surface of the nano-magnetic particles; then under the action of an external magnetic field, let it stand for 2 minutes, the magnetic particles settle down, remove the supernatant, add 500 μL of cleaning solution, remove the magnetic field, shake to make Magnetic particle resuspension; repeat this washing step 3 times to remove unbound biotin-labeled osteocalcin to obtain the first solution;

[0060] (2) Add 60 μL of horseradish peroxidase-labeled osteocalcin antibody solution to the first solution, mix well, and incubate at 37°C for 15 minutes; then, under the action ...

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Abstract

The invention provides a kit for detecting osteocalcin. The kit is high in detecting sensitivity and good in reliability. The kit for detecting the osteocalcin is characterized in that the kit comprises a calibration product, a quality control product, a biotin-labeled osteocalcin antibody solution, a horseradish peroxidase-labeled osteocalcin antibody solution, a streptavidin-labeled nano magnetic particle suspension and a substrate solution; the calibration product comprises an osteocalcin antigen solution with the concentration being 0 ng/mL, 10 ng/mL, 25 ng/mL, 50 ng/mL, 75 ng/mL and 250 ng/mL; the concentration of the biotin-labeled osteocalcin antibody solution is 1.0 [mu]g/mL; the concentration of the streptavidin-labeled nano magnetic particle suspension is 0.5 mg/mL; and the concentration of the horseradish peroxidase-labeled osteocalcin antibody solution is 1.0 [mu]g/mL.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis of biological immunity of medical equipment, and in particular relates to a kit for detecting osteocalcin and a use method. Background technique [0002] Osteoporosis is a static disease: some non-specific symptoms before fractures are often ignored: such as low back pain. Most cases are diagnosed after fractures, and the associated complications are mostly fractures, which greatly increases medical costs. Early diagnosis can help reduce healthcare costs. More and more hospital departments have opened osteoporosis detection, such as: endocrinology, orthopedics, obstetrics and gynecology, traditional Chinese medicine, etc., and gradually began to pay attention to the admission and treatment of osteoporosis patients. At present, hospitals at all levels in my country mainly use bone mineral density as an auxiliary diagnostic standard, and have not fully used bone metabolism markers. [0003] Oste...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/54346G01N33/581G01N33/68G01N2800/108
Inventor 韩霜白仲虎张俊兰
Owner JIANGNAN UNIV
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