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PIVKA-II (protein induced by vitamin K absence-II) magnetic particle chemiluminescence immune assay determination kit and preparation method thereof

A technology for abnormal prothrombin and chemiluminescence immunity, which is applied in the field of immunoassay medical analysis, can solve problems such as unsatisfactory results, and achieve the effect of increasing the effective coating amount, high sensitivity, and easy promotion and application

Pending Publication Date: 2019-03-05
江苏麦得科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented method allows us to combine powerful chemical reactions that produce very small amounts of energy into an assay system without losing its ability or affecting other important components like fluorescein labelled probes. By combining these techniques together we have developed a highly sensitive and selective way to measure blood clotting factors (procoagulants) accurately overcomes problems associated with current laboratory tests such as subjectivity and lack of accuracy due to variations between individuals' responses during testing procedures.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the accuracy and reliability of detecting certain types of tissue from noncancerous samples like those found during routine medical examinsations without causing damage due to their presence.

Method used

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  • PIVKA-II (protein induced by vitamin K absence-II) magnetic particle chemiluminescence immune assay determination kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Implementation example 1 Magnetic particles coated with anti-fluorescein isothiocyanate (FITC) monoclonal antibody

[0020] Activate the magnetic particles with a particle size of 0.1-5μm with glutaraldehyde, stir at room temperature, mix for 2 hours, apply a magnetic field, let stand for 20-25min, pour out the supernatant, and use 0.01mol / L phosphoric acid with a pH value of 7.4 Wash three times with salt buffer solution and suspend with the solution at a concentration of 50-100 mg / mL; add 60-100 μg of anti-FITC monoclonal antibody to each ml of suspension, stir overnight at 4°C, add a magnetic field, and let stand for 10- After 15 minutes, pour out the supernatant, and block with phosphate buffer (pH 7.2) containing 0.2%-1.0% bovine serum albumin, 0.5%-1.0% casein, and 0.02mol / L at room temperature for 3-4 hours; finally Wash 3-5 times with phosphate washing buffer solution with a pH value of 7.4, containing Tween-20 and sodium azide preservative, and use this solutio...

Embodiment 2

[0021] Implementation example 2 washing buffer

[0022] Wash buffer is phosphate buffered saline containing 0.2-0.5% Tween-20, 0.5mL / L Proclin 300 preservative, pH 7.4. Dilute 10 times with distilled water before use.

Embodiment 3

[0023] Implementation Example 3 The usage method of this kit

[0024] 1. Sample pretreatment

[0025] The fasting morning serum samples were taken from people, centrifuged at 3000rpm for 5min, and the upper serum was taken for analysis.

[0026] 2. Detection method

[0027] Abnormal prothrombin (PIVKA-Ⅱ) magnetic particle chemiluminescence immunoassay assay kit.

[0028] Before using this kit for experiments, it is necessary to take out the magnetic particle solution, FITC-labeled PIVKA-II antibody, standard substance, quality control substance, chemiluminescence substrate and other experimental reagents and place them at room temperature for 15-30 minutes, and then equilibrate to room temperature; Secondly, adjust the water bath or thermostat to 37°C for convenience; after that, prepare a suitable micro-sampler and corresponding tips, and check whether the chemiluminescence instrument is working normally.

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Abstract

The invention belongs to the technical field of immune assay medical detection and particularly provides a PIVKA-II (protein induced by vitamin K absence-II) magnetic particle chemiluminescence immuneassay determination kit and a preparation method thereof. The kit mainly comprises (1) a PIVKA-II standard product; (2) anti-FITC (fluorescein isothiocyanate) monoclonal antibody coated magnetic particles; (3) an alkaline phosphatase marked PIVKA-II sample and fluorescein isothiocyanate marked PIVKA-II; (4) an alkaline phosphatase marked chemiluminescent substrate; (5) subpackage of the standardproduct, magnetic particle solution, mixed solution, sample dilution liquid, washing liquid, the chemiluminescent substrate and reaction tubes. By combination of the enzyme marking technique, the magnetic particle separation technique and the chemiluminescence detection technique, an approximately homogenous reaction system is provided, and simplicity, sensitivity, high specificity, wide linear range, high repeatability and the like are realized.

Description

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Claims

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Application Information

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Owner 江苏麦得科生物科技有限公司
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