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Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III), and preparation method of kit

A chemiluminescence immunoassay and quantitative detection technology, applied in the field of immunoassay medicine, can solve the problems of short validity period of reagents, narrow detection range, low sensitivity, etc., and achieve the effects of shortened reaction time, easy operation and high sensitivity

Active Publication Date: 2014-12-10
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The problem to be solved by the present invention is to provide PCⅢ magnetic particle chemiluminescence immunoquantitative detection kit and its preparation method, which avoids the shortcomings of radioimmunoassay reagents such as short validity period, radioactive pollution, cumbersome operation, etc., and solves the problem of low sensitivity and detection Narrow scope, high cost defects

Method used

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  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III), and preparation method of kit
  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III), and preparation method of kit
  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III), and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of Type Ⅲ Procollagen (PCⅢ) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit Ⅰ

[0038] (1) Preparation of PCⅢ calibrator: Add PCⅢ pure product to physiological saline solution containing 5% bovine serum, the concentrations are 0, 20, 50, 100, 200, 500 ng / mL respectively.

[0039] (2) Preparation of PCⅢ quality control products:

[0040] The PCⅢ antigen was prepared with a normal saline solution containing 5% bovine serum to prepare a low-value quality control product and a high-quality control product. The concentration of QcL was 42.5 ng / mL, and the concentration of QcH was 255 ng / mL.

[0041] (3) Preparation of magnetic particle-streptavidin suspension

[0042] Take 100mL of 0.1mol / L 2-morpholineethanesulfonic acid solution (MES solution), add 10mg of magnetic particles and 3mg of streptavidin (SA) in turn, stir for 30min, and then add 10mg / mL of EDC solution (1-B Base-(3-dimethylaminopropyl, also known as: carbodiimide hydrochlo...

Embodiment 2

[0067] Example 2: Preparation of Type Ⅲ Procollagen (PCⅢ) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit Ⅱ

[0068] (1) Preparation of PCⅢ calibrator: Add PCⅢ pure product to physiological saline solution containing 5% bovine serum, the concentrations are 0, 20, 50, 100, 200, 500 ng / mL respectively.

[0069] (2) Preparation of PCⅢ quality control products:

[0070] The PCⅢ antigen was prepared with a normal saline solution containing 5% bovine serum to prepare a low-value quality control product and a high-quality control product. The concentration of QcL was 57.5 ng / mL, and the concentration of QcH was 345 ng / mL.

[0071] (3) Preparation of magnetic particle-streptavidin suspension

[0072] Take 100mL of 0.1mol / L 2-morpholineethanesulfonic acid solution (MES solution), add 10mg of magnetic particles and 3mg of streptavidin (SA) in turn, stir for 30min, and then add 10mg / mL of EDC solution (1-B Base-(3-dimethylaminopropyl, also known as: carbodiimide hydroch...

Embodiment 3

[0086] Example 3: Preparation of Type Ⅲ Procollagen (PCⅢ) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit Ⅲ

[0087] (1) Preparation of PCⅢ calibrator: Add PCⅢ pure product to physiological saline solution containing 5% bovine serum, the concentrations are 0, 20, 50, 100, 200, 500 ng / mL respectively.

[0088] (2) Preparation of PCⅢ quality control products:

[0089] The PCⅢ antigen was prepared with a normal saline solution containing 5% bovine serum to prepare a low-value quality control product and a high-quality control product. The concentration of QcL was prepared between 50.2ng / mL and the concentration of QcH was prepared at 305ng / mL.

[0090] (3) Preparation of magnetic particle-streptavidin suspension

[0091] Take 100mL of 0.1mol / L 2-morpholineethanesulfonic acid solution (MES solution), add 10mg of magnetic particles and 3mg of streptavidin (SA) in turn, stir for 30min, and then add 10mg / mL of EDC solution (1-B Base-(3-dimethylaminopropyl, also know...

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Abstract

The invention discloses a quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III). The kit comprises PC III calibrators, magnetic particle suspension coupled with streptavidin, a PC III antibody labeled with biotin, a PC III abzyme combination, a PC III quality controller, chemiluminescence liquor A, chemiluminescence liquor B, 20 times concentrated washing liquor, and a reaction tube, wherein the concentrations of the PC III calibrators include 0, 20ng / ml, 50ng / ml, 100ng / ml, 200ng / ml and 500ng / ml, and diluent of the calibrators is a physiological saline solution containing 5% of bovine serum; and enzyme adopted by the PC III abzyme combination is horse radish peroxidase with the purity RZ being more than or equal to 3.0 and the activity being more than or equal to 250U / ml. The invention also discloses a preparation method of the kit. Compared with the conventional kit, the quantitative detection kit is simple and convenient to operate, is safe, does not cause environment pollution, and also has the advantages of wide concentration range, low cost, good stability and the like of detection samples.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a type III procollagen (PCIII) magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] There are five types of collagen (types I, II, III, IV, and V) in tissues. Normal liver mainly contains type Ⅰ, Ⅲ, Ⅳ, Ⅴ procollagen, which is the main collagen in the hyperplasia of liver extracellular interstitial connective tissue. PCⅢ is synthesized in the cells and then secreted into the blood. PⅢP is the amino-terminal peptide fragment after degradation of PCⅢ, which is only 1 / 10 of PCⅢ, with a molecular weight of 45,000. When liver fibrosis occurs, the synthesis of PCⅢ by fibroblasts increases and the degradation decreases. In chronic liver and cirrhosis, liver fiber synthesis and metabolism are vigorous, and serum PCⅢ increases. Therefore, when there is active fibrosis, serum PCⅢ increa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
Inventor 刘萍范利花
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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