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Creatine kinase isozyme detection kit and preparation thereof

A creatine kinase and kit technology, applied in the field of creatine kinase isoenzyme detection kit and its preparation, can solve problems such as unsatisfactory, inappropriate, false negative, etc., achieve large market competitiveness, prolong service life, The effect of reducing manufacturing costs

Active Publication Date: 2014-07-16
BEIJING JIUJIAYI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 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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  • Creatine kinase isozyme detection kit and preparation thereof
  • Creatine kinase isozyme detection kit and preparation thereof
  • Creatine kinase isozyme detection kit and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 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.

[0034] The specific process is as follows:

[0035] 1) The preparation of colloidal gold of 50nm is as follows:

[0036] Put a magnetic stirrer in a cleaned 1000ml round bottom flask, and add 500ml of ultrapure water;

[0037] 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;

[0038]Turn off the heating switch and continue to stir for 10 minutes, then stop stirring and cool to room temperatu...

Embodiment 2

[0057] 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.

[0058] 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

[0060] 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.

[0061] 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 creatine kinase isozyme detection kit. The kit is based on colloidal gold immunoturbidimetry, and comprises reagents R2, and the reagents R2 are solutions of gold nanoparticles marked with creatine kinase isozyme antibodies. The kit is characterized in that the particle diameter of the gold nanoparticles is 62.2 nm to 79.1 nm, and the mass ratio of the gold nanoparticles and the antibodies is 50:20-60. The invention further provides a preparation method of the kit. The kit has the advantages of being high in sensitivity, strong in specificity, fast in response and good in stability, no sediment is generated after a 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 a creatine kinase isoenzyme detection kit based on colloidal gold immunoturbidimetry and its preparation. Background technique [0002] Creatine kinase (CK) has three major isozymes, namely CK-MM, CK-MB, and CK-BB. Creatine kinase MB isoenzyme (CK-MB), mainly present in the myocardium, is the most specific enzyme in the myocardial enzyme spectrum. Since the 1980s, it has been recognized as the "gold standard" for myocardial enzyme diagnosis. Early diagnosis of acute myocardial infarction (AMI) is a sensitive and specific indicator for diagnosis and monitoring of AMI patients. [0003] Immunosuppression is the most commonly used clinically. The principle is: the anti-CK-M antibody in the immunosuppression reagent inhibits the CK-M subunit, that is, inhibits the activity of CK-MM and the activity of the CK-M subunit of CK-MB. Measure the activity of the uninhibited CK-B monomer in the sample (equivalent to half the activity o...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/532
CPCG01N33/54346G01N33/54393G01N33/573
Inventor 侯淑霞王万霞
Owner BEIJING JIUJIAYI TECH
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