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Kit for detecting alpha fetal protein and preparation method of kit

A detection kit and alpha-fetoprotein technology, applied in the field of alpha-fetoprotein detection kits, can solve problems such as inappropriateness, false negatives, unsatisfaction, etc., and achieve the effects of prolonging service life, reducing manufacturing costs, and increasing market competitiveness

Active Publication Date: 2014-07-30
BEIJING JIUJIAYI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The research of the present invention shows that many nanoparticles do not meet the requirements of the test within this range, such as: when the magnetic nanoparticles such as iron oxide are more than 40nm, they are easy to coagulate and settle due to factors such as paramagnetism and specific gravity, and cannot achieve proper stability; The stability of gold nanoparticles in the range of 80-150nm is too poor and unsuitable, etc.
According to the teaching of Chinese patent CN102749454, the present invention tests colloidal gold particles with a diameter of 35nm-60nm, and finds that the sensitivity is poor and cannot meet the clinical requirements for sensitivity less than 1ng / mL and the minimum detection limit of 5ng / mL , and the linear range does not cover the concentration range of normal physiological and pathological conditions in clinical practice, so false negative results are often caused; at the same time, there is also a long reaction time (equilibrium time 5-10min), which cannot meet the requirements of rapid testing

Method used

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  • Kit for detecting alpha fetal protein and preparation method of kit
  • Kit for detecting alpha fetal protein and preparation method of kit
  • Kit for detecting alpha fetal protein and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of reagent R2 in this example is divided into three steps. Firstly, a colloidal gold solution is prepared, then the antibody is coupled to the gold nanoparticles, and finally it is mixed with buffer salt, coagulant and preservative to form a complete system.

[0033] The specific process is as follows:

[0034] 1) The preparation of colloidal gold of 50nm is as follows:

[0035] Put a magnetic stirrer in a cleaned 1000ml round bottom flask, and add 500ml of ultrapure water;

[0036] Add 5ml concentration and be 1% (w / v) chloroauric acid, stir at about 600rpm at a high speed, and be heated to solution boiling, be that 1% (w / v) sodium citrate solution is added in the flask rapidly then with 6ml concentration, keep Heating and stirring vigorously for 10 minutes, the color of the solution first turns black, and then gradually turns purple;

[0037] Turn off the heating switch and continue to stir for 10 minutes, then stop stirring and cool to room temperat...

Embodiment 2

[0056] The only difference between Example 2 and Example 1 is that the particle size of the colloidal gold particles prepared in Example 2 is 55 nm by adjusting the amount of reducing agent added.

[0057] Prepare 55nm particles and add reducing agent 1% sodium citrate to 5.6ml. The obtained colloidal gold particles were observed by transmission electron microscope, and the diameter was about 56.2±1.5nm.

Embodiment 3

[0059] The only difference between Example 3 and Example 1 is that the particle size of the colloidal gold particles prepared in Example 3 is 62.2 nm by adjusting the amount of reducing agent added.

[0060] Prepare 62.2nm particles and add reducing agent 1% sodium citrate to make 5.4ml. The obtained colloidal gold particles were observed by transmission electron microscope, and the diameter was about 62.2±1.1nm.

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Abstract

The invention provides a kit for detecting an alpha fetal protein. The kit is operated based on a colloidal gold immunoturbidimetry and comprises a reagent R2, wherein the Reagent R2 is a solution which contains gold nanoparticles marked with alpha fetal protein antibodies. The kit is characterized in that the particle diameters of the contains gold nanoparticles are 62.2-79.1 nomameters; the mass ratio of the contains gold nanoparticles to the alpha fetal protein antibodies is 50:(20-60). The invention also provides a preparation method of the kit. The kit provided by the invention has the characteristics of high sensitivity, high specificity, fastness in reaction and good stability, precipitations can not be generated after reaction, a biochemical analyzer is convenient to clean and the service life of the biochemical analyzer is prolonged.

Description

technical field [0001] The application relates to an alpha-fetoprotein detection kit based on colloidal gold immunoturbidimetry and its preparation. Background technique [0002] As early as 1944, the existence of alpha-fetoprotein had been studied, but it was not until 1965 that American scientists Bergstyandh and Czar observed that the serum of two patients with primary liver cancer could react with the monovalent antiserum of anti-fetal serum, indicating that the original There is a common special component in the serum of patients with primary liver cancer and human fetal serum, and this special protein component is called α-fetoprotein (AFP). Subsequent research has found that the protein cross-reacts between 18 mammalian species. For a long time, people have only used AFP as a tumor marker and one of the diagnostic indicators for primary liver cancer (HCC). [0003] Due to the time-consuming and complicated operation of enzyme-linked immunosorbent assay, it can no lo...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/54346G01N33/6803G01N2800/7028
Inventor 侯淑霞王万霞
Owner BEIJING JIUJIAYI TECH
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