Method for determining gibberellin based on hybridization chain-reaction signal amplification technology

A technology of hybridization chain reaction and signal amplification, which is applied in the direction of chemical reaction of materials for analysis, biological testing, material inspection products, etc., and can solve the problems of cumbersome pretreatment methods and low sensitivity

Active Publication Date: 2014-03-26
凯惠睿智生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Some pretreatment methods in these methods ...

Method used

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  • Method for determining gibberellin based on hybridization chain-reaction signal amplification technology
  • Method for determining gibberellin based on hybridization chain-reaction signal amplification technology
  • Method for determining gibberellin based on hybridization chain-reaction signal amplification technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Effect of the amount of antibody-modified magnetic beads on the intensity of chemiluminescence

[0055] According to the technical scheme, the test of the influence of the amount of antibody-modified magnetic beads on the intensity of chemiluminescence was carried out. The test results are as follows: figure 2 As shown in (A), the intensity of chemiluminescence increases gradually when the amount of antibody-modified magnetic beads increases from 2 μL to 10 μL. If the amount of antibody-modified magnetic beads is increased, the intensity of chemiluminescence decreases slowly, so 10 μL of antibody-modified magnetic beads is preferably used in the present invention.

Embodiment 2

[0056] Example 2 Effect of the amount of antibody and DNA1 modified colloidal gold on the intensity of chemiluminescence

[0057] The effect of the amount of antibody and DNA1-modified colloidal gold on the chemiluminescence intensity was tested. When the amount of antibody-modified magnetic beads was kept at 10 μL, when the amount of antibody and DNA1-modified colloidal gold was increased from 5 μL to 15 μL, the chemiluminescence intensity continued to increase . Continue to increase the amount of antibody and DNA1-modified colloidal gold, the chemiluminescence intensity gradually increases, and finally reaches the maximum at 20 μL, and then remains at this level, almost unchanged, the test results are as follows figure 2 (B). This is because the reaction was saturated when the amount of antibody and DNA1-modified colloidal gold was 20 μL. Therefore, 20 μL is the optimum amount of antibody and DNA1 modified colloidal gold. Therefore, the dosage of the antibody of the pres...

Embodiment 3

[0058] Example 3 Effect of reaction time between standard solution and antibody-labeled magnetic beads on chemiluminescent intensity

[0059] When the reaction between the standard solution and the antibody-labeled magnetic beads changed from 5 minutes to 30 minutes, the chemiluminescent intensity increased sharply, and then tended to be stable. The results are as follows: figure 2 (C) shown. Continuing to increase the reaction time does not cause a significant increase in the intensity of chemiluminescence. Therefore, the reaction time between the sample of the present invention and the antibody-labeled magnetic beads is preferably 30 minutes.

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Abstract

A disclosed method for determining gibberellin based on hybridization chain-reaction signal amplification technology is characterized by: fixing gibberellin antibody on carboxylated magnetic beads so as to form a sandwich-type immunization complex by gibberellin antibody on the magnetic beads, gibberellin antibody on colloidal gold and gibberellin when the target gibberellin exists, adding H1 and H2 both rich in G base, exciting a hybridization chain reaction by DNA1 on colloid gold so as to form a DNA long chain rich in G base on the immunization complex and generate chemical luminescence under the effect of a chemiluminescence agent 3,4,5-trimethoxyphenylglyoxal, and determining gibberellin according to chemiluminescence. The method has the characteristics of high sensitivity, high selectivity, low cost, simple operation and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for measuring gibberellin based on hybridization chain reaction signal amplification technology. Background technique [0002] Gibberellins (GA, 2,4α,7-trihydroxy-1-methyl-8-methylenegibber-3-ene-1,10-dicarboxylate-1,4α-lactone) are ubiquitous A phytohormone in higher plants, involved in regulating various physiological processes of plant growth and development, such as promoting cell division, plant dwarfing, etc. At present, there is no regulation on the maximum residue limit of gibberellin in my country, but the United States, Japan and other countries stipulate that the maximum residue limit of gibberellin in fruits and vegetables is 0.2mg / kg (Zhao Yingbo, Zhou Yanming, Xin Xue, Wang Yansong. Food Science , 2011, 32(06): 209; Lu Baoying. Encyclopedia of Food Analysis. Beijing: Higher Education Press, 1997:723). The methods for determining gibberellins mainl...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/74
Inventor 混旭苏国庆
Owner 凯惠睿智生物科技(上海)有限公司
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