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Detection kit for urine beta-collagen degradation products and preparation method thereof

A technology for detecting kits and degradation products, applied in the field of medical devices, can solve the problems of lack of specificity and good sensitivity

Inactive Publication Date: 2012-08-29
GUANGZHOU GUCON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is a lack of relevant detection kits with specificity, good sensitivity and simple operation on the market

Method used

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  • Detection kit for urine beta-collagen degradation products and preparation method thereof
  • Detection kit for urine beta-collagen degradation products and preparation method thereof
  • Detection kit for urine beta-collagen degradation products and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1), the main components of the urinary β-collagen degradation product detection kit described in the present embodiment are as follows:

[0024] 1. Microplate

[0025] 6-strip microwell plates (2x 8 wells / strip) pre-coated with CTX-I antigen, placed in a plastic frame.

[0026] 2. CTX-I Standard 0

[0027] One bottle (11 mL) of ready-to-use buffer solution.

[0028] 3. CTX-I standard 1-5

[0029] Five bottles (0.3mL / bottle) of CTX-I standard that can be used directly.

[0030] 4. Urine quality control

[0031] Two bottles (0.5mL / bottle) of human urine that can be used directly.

[0032] 5. Primary antibody solution

[0033] One bottle (12mL) of ready-to-use rabbit anti-CTX-I antibody solution.

[0034] 6. Peroxidase-conjugated antibody

[0035] One bottle (12mL) of ready-to-use horseradish peroxidase-conjugated anti-rabbit antibody at a working concentration of 1:3000.

[0036] 7. Substrate solution

[0037] One bottle (12 mL) of ready-to-use tetramethylbenzi...

Embodiment 2

[0079] Example 2: Using the detection kit described in Example 1 to detect the sample

[0080] All solutions were equilibrated to room temperature before use. Experiments were carried out at 18-22°C.

[0081] Determine the number of microplates needed for the experiment. In addition, a total of 16 wells are required for each round of experiments for standards and quality controls. Place the appropriate number of microplates on the plastic frame. Seal unused microplates in a foil pouch with desiccant.

[0082] 1. Add standards, quality controls and samples to be tested

[0083] Pipette 15 μl of standard, quality control or sample to be tested in a microwell plate.

[0084] 2. Microplate Incubation

[0085] Add 100 μl of primary antibody solution to each well. Seal the microplate with Parafilm and incubate for 60±5 min at 18-22°C on a microplate shaker at 300 rpm.

[0086] 3. Rinse

[0087] The rinse solution 50x was diluted in the ratio of 1 volume of concentrated buff...

Embodiment 3

[0123] Precision Analysis

[0124] Using the urinary β-collagen degradation product detection kit described in Example 1, and according to the detection method described in Example 2, the accuracy was analyzed.

[0125] The following results were obtained by testing four samples, each sample was measured in quintuplicate, and measured continuously for five days (each sample was measured 25 times in total).

[0126] Inter-assay CV (n=25)

[0127]

[0128] Within-assay CV (n=25)

[0129]

[0130] Dilution / Linearity

[0131] The linear range of this kit is from 50μg / L to 6750μg / L.

[0132] The urine sample with a CTX-I concentration of 3450-4340 μg / L was diluted with a standard 0, and then measured using the kit described in Example 1.

[0133] The concentration of the original urine sample was set at 100%.

[0134]

[0135] Recovery rate

[0136] Mixed samples were prepared by adding different amounts of CTX-I antigen to three different urine samples.

[0137] ...

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Abstract

The invention discloses a detection kit for urine beta-collagen degradation products. The detection kit mainly comprises microwell plates coated with CTX-I antigens and a rabbit anti CTX antibody solution, wherein the amount of the coated CTX-I antigens of each microwell plate is 0.48-7.2 mug, and the valence of rabbit anti CTX antibodies achieves 1:10000. The detection kit for the urine beta-collagen degradation products has the advantage of good specificity, sensitivity and precision.

Description

technical field [0001] The invention belongs to the field of medical devices, and in particular relates to a urine β-collagen degradation product detection kit and a preparation method thereof. Background technique [0002] Type I collagen accounts for more than 90% of the organic matrix of bone and is mainly synthesized in bone. During bone renewal, type I collagen is degraded and short peptide fragments enter the urine. These fragments can be detected by urine β-collagen degradation product detection kit, and it has been reported in the literature that this analysis is helpful for the research of patients suffering from bone metabolic diseases after anti-resorptive therapy. [0003] At present, there is a lack of relevant detection kits with specificity, good sensitivity, and simple operation on the market. Contents of the invention [0004] One of the objectives of the present invention is to provide a detection kit for urinary β-collagen degradation products. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/532
Inventor 郭志程
Owner GUANGZHOU GUCON BIOTECH
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