HBP magnetic particle chemiluminiscence method detection kit and preparation method thereof

A detection kit and magnetic particle technology, which is applied in the field of heparin-binding protein detection, can solve the problems that the accuracy of the test cannot be guaranteed, the clinical judgment basis cannot be provided, and interferences are removed, so as to reduce the steric hindrance effect and improve the detection sensitivity. and specificity, reducing the effect of non-specific reactions

Active Publication Date: 2020-09-18
SUZHOU KANGHESHUN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although colloidal gold method and immunofluorescence chromatography can be quickly detected, they can only perform qualitative or semi-quantitative detection, with low accuracy and cannot provide accurate clinical judgment basis
At the same time, limited by the method itself, due to the use of a heterogeneous reaction system, the detection precision is poor, and the interference cannot be removed from the operating procedure, so that the accuracy of the test cannot be guaranteed

Method used

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  • HBP magnetic particle chemiluminiscence method detection kit and preparation method thereof
  • HBP magnetic particle chemiluminiscence method detection kit and preparation method thereof
  • HBP magnetic particle chemiluminiscence method detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Preparation of biotin-labeled HBP capture antibody solution

[0028] Dissolve biotin in dimethyl sulfoxide to 1mg / ml in advance, mix biotin-dimethyl sulfoxide solution and HBP capture antibody at a mass ratio of 1:10, control the volume within 100ul, add carbonate buffer The volume of the solution was made up to 150ul, and incubated at 37°C for 1 hour. After the reaction, the solution was replaced by ultrafiltration using a 50kDa ultrafiltration tube, and the ultrafiltrate was replaced 3 times during the reaction. The final antibody concentrate was collected in 50mM pH7.3 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 0.5% lauryl polyoxyethylene ether, 50mM potassium chloride, 2% sucrose, 4% arginine, 0.2 In % Proclin-300, the final concentration of biotinylated HBP capture antibody was 2 μg / mL.

[0029] (2) Preparation of biotin-labeled bovine serum albumin solution

[0030] Dissolve biotin in dimethyl sulfoxide to 1mg / ml in advance, mix biotin-dimethyl sul...

Embodiment 2

[0048] Analytical sensitivity of reagents (blank limit)

[0049] Using the reagents in Example 1 (4) and (5), select physiological saline as a blank sample test, repeat the test 20 times, test with a semi-automatic chemiluminescence immunoassay analyzer, and record the luminescence value (RLU). The specific data are shown in Table 2 . Calculate the mean value (X) and standard deviation (SD) of 20 test results, bring the calculated value of X±2SD into the standard curve established in Example 1 to calculate the concentration value, and the obtained concentration value is used as the analytical sensitivity of the reagent (blank limit ). The test results are shown in Table 2, the analytical sensitivity (blank limit) of the reagent in (4) is 0.34ng / mL, the analytical sensitivity (blank limit) of the reagent in (5) is 2.11ng / mL, (4 The analytical sensitivity of the reagents in (5) is much lower than that of the reagents in (5), so the reagents in (4) are selected to complete the ...

Embodiment 3

[0052] The specificity of embodiment 3 reagents

[0053] Using the reagents in Example 1 (4) and (5), add calibrator dilution (blank group) and calibration Different concentrations of specific interfering substances bilirubin (D experimental group), triglyceride (G experimental group), total protein (Z experimental group) and rheumatoid factor (L experimental group) were dissolved in the product diluent. The theoretical value reductions of the low-value quality control and high-value quality control in the group were 6.4 and 32 ng / mL. By comparing the test results of the blank group and the experimental group, the smaller the relative deviation between the average value of the five tests and the theoretical value, the better the specificity.

[0054] Table 3 Specificity detection results of Example 1 (4)

[0055]

[0056] Table 4 Specificity detection results of Example 1 (5)

[0057]

[0058] The relative deviation of the five detection results in Example 1 (4) is mu...

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Abstract

The invention discloses a magnetic particle chemiluminiscence detection kit for heparin-binding protein, a preparation method of the magnetic particle chemiluminiscence detection kit and application of the magnetic particle chemiluminiscence detection kit to immunological detection of the heparin-binding protein. The magnetic particle chemiluminiscence detection kit comprises a streptavidin-coatedmagnetic particle suspension solution, a biotin-labeled heparin-binding protein capture antibody and biotin-labeled bovine serum albumin mixed solution, a luminous marker coupled heparin-binding protein detection antibody solution, a heparin-binding protein series calibration product and a quality control product. The kit provided by the invention utilizes the advantages of a magnetic particle chemiluminescence detection method, and has the characteristics of simple operation, strong specificity, high sensitivity, wide linear range and the like when a full-automatic chemiluminescence immunoassay analyzer is used for detecting the heparin-binding protein in a specimen.

Description

technical field [0001] The invention belongs to the field of medical immunodiagnostic reagents, in particular to the detection of heparin-binding protein based on chemiluminescence. Background technique [0002] Heparin Binding Protein (HBP for short) is a granular protein secreted by neutrophils, which has a bactericidal effect and is also involved in the regulation of inflammatory responses and blood coagulation processes. In 1984, Shafer et al. first discovered and isolated this protein, and named it CAP37 (Cationic Antimicrobial Protein, molecular weight 37kDa) according to its electropositive characteristics and bactericidal function. Later, azurocidin and heparin-binding protein (HBP-Heparin Binding Protein), which has a strong binding ability to heparin, were isolated from the azurophilic blue granules in polymorphonuclear leukocytes. Further studies on protein structure and gene sequencing show that these three proteins are actually the same protein, which is repres...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/577G01N33/6893G01N2333/47G01N2800/7095
Inventor 王明陈胜胜刘向晖汪春芳
Owner SUZHOU KANGHESHUN MEDICAL TECH
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