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Quantitative determination RBP4 kit by chemiluminescence magnetic enzymoimmune method

A chemiluminescence and reagent kit technology, which is applied in the field of chemiluminescence magnetic enzyme immunoassay, can solve the problems of radioactive pollution and short half-life, and achieve the effects of high specificity, fast operation and detection, and stable luminescence performance

Active Publication Date: 2009-06-10
WUHAN EASYDIAGNOSIS BIOMEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While inheriting the advantages of high sensitivity of radioimmunoassay, it overcomes the shortcomings of radioactive pollution and short half-life

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Preparation of the kit:

[0034] a.RPB4 immunomagnetic beads preparation: select superparamagnetic polymer magnetic microspheres as the carrier of magnetic beads (trade name Bruker, manufactured by BioSciences Corporation), and superparamagnetic ferriferrous oxide with an average diameter of about 8-10 nanometers is dispersed inside it ; The amino-modified activated magnetic beads were washed with PBS buffer solution, then added to 5% glutaraldehyde / PBS solution at room temperature, protected from light and oscillated for magnetic separation, and the activated magnetic beads were prepared after washing for many times; the activated magnetic beads were taken The beads and the specific mouse anti-human RBP4 antibody I were incubated at room temperature at a ratio of 100ug antibody / mg magnetic beads. After magnetic separation, an excess of 10% BSA solution was added to block the free aldehyde groups of the unbound antibody, and the specific mouse anti- Immunoactive magn...

Embodiment 2

[0043] Using the same kit as in Example 1, the serum samples to be tested were determined.

[0044] Determination of RBP4 concentration

[0045] Serum sample 2 to be tested (26-year-old healthy female). Serum samples to be tested are diluted to become samples to be tested. Take 20ul of the sample, add 100ul of specific RBP4 immunomagnetic beads, incubate at room temperature for 20min to bind the RBP4 antigen to be tested in the sample, and then add alkaline phosphatase to label specific mouse anti-human RBP4 Antibody 20ul, incubated at room temperature for 20min, alkaline phosphatase-labeled specific mouse anti-human RBP4 antibody combined with the antigen to be tested, using magnetic microbeads as a solid phase carrier to form a RBP4 double-antibody sandwich complex on its surface. After magnetic separation and washing, 10ul / ml AMPPD / carbonate solution was added to the system for 30 minutes, and the alkaline phosphatase in the double-antibody sandwich complex acted on the lu...

Embodiment 3

[0048] Using the same kit as in Example 1, the serum samples to be tested were assayed.

[0049] Determination of RBP4 concentration

[0050] Serum sample 3 to be tested (60-year-old male, overweight, with insulin resistance, but has not yet reached the diagnostic criteria for type 2 diabetes). Serum samples to be tested are diluted to become samples to be tested. Take 20ul of the sample, add 100ul of specific RBP4 immunomagnetic beads, incubate at room temperature for 20min to bind the RBP4 antigen to be tested in the sample, and then add alkaline phosphatase to label specific mouse anti-human RBP4 Antibody 20ul, incubated at room temperature for 20min, alkaline phosphatase-labeled specific mouse anti-human RBP4 antibody combined with the antigen to be tested, using magnetic microbeads as a solid phase carrier to form a RBP4 double-antibody sandwich complex on its surface. After magnetic separation and washing, 10ul / ml AMPPD / carbonate solution was added to the system for 30 ...

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Abstract

The invention relates to a medical testing kit for performing quantitative detection on human serum RBP4 using chemiluminescence magnetic-enzyme immunotherapy. The kit is composed of four reagent parts: specificity mouse anti-human RBP4 custodite immunomagnetic beads, enzyme labelling specificity mouse anti-human RBP4 antibody II, chemiluminescence substrate, corresponding titer and quality control liquid. The using method of the kit comprises: using bead particulates as solid phase carrier, combining specificity mouse anti-human RBP4 antibody I on the surface, forming RBP4 specificity immunocompetence beads, capturing antigen RBP4 to be detected in the enzyme labelling specificity mouse anti-human RBP4 antibody II, forming double antibody sandwich composite on the surface of the beads, wherein enzyme marked on the composite reacts with corresponding irradiance substrate in the reaction system to form stable luminous signals, thereby reaching quantitative detection and analysis on RBP4 through strength of the detection light signals. The invention has the advantages of high sensitivity, high specificity, simple and fast operation.

Description

technical field [0001] The invention relates to a chemiluminescent magnetic enzyme immunoassay technique, in particular to a kit for quantitatively detecting serum retinol binding protein 4 (RBP4) by a chemiluminescent magnetic enzyme immunoassay. Background technique [0002] With the improvement of people's living standards and changes in lifestyle, type 2 diabetes has become a chronic disease that seriously endangers human health. In the past 20 years, the incidence of diabetes has increased worldwide. It is estimated that the number of people with diabetes in the world will increase to about 220 million in 2010. The number of people with prediabetes, including impaired glucose tolerance and impaired fasting blood sugar, will also reach or even exceed the number of people with diabetes. The treatment of type 2 diabetes and its complications has become a heavy burden. Therefore, it is of great significance for its early diagnosis and active prevention. Insulin resistance ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/546G01N21/76
Inventor 王颖陈莉莉
Owner WUHAN EASYDIAGNOSIS BIOMEDICINE
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