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Luminol direct bonded nano gold nucleic acid analyzing probe and application thereof

A technology of nucleic acid analysis and direct bonding, which is applied in the direction of chemical reaction of materials for analysis, measurement/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problem of unsatisfactory sensor stability due to the lack of research on the application of nano-gold , complex condensation reaction process, etc., to achieve excellent labeling properties, good biocompatibility, and simple labeling methods

Inactive Publication Date: 2011-04-20
UNIV OF SCI & TECH OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the complexity of the condensation reaction process and the need to purify and separate the products after the reaction, resulting in high analysis costs
(2) Enzymes, such as DNase, alkaline phosphatase, peroxidase, digoxin, etc., mostly use enzymes to catalyze specific chemiluminescent reactions, but due to the chemical modification of enzymes, their reactivity is somewhat different. Therefore, there are problems of easy inactivation and poor analysis stability
But measuring H 2 o 2 The sensitivity is low, and its detection limit is only 1×10 -7 mol / LH 2 o 2
In addition, the stability of the sensor is not ideal, so it is difficult to be used for the determination of actual samples
In this work, the application of luminol directly bonded gold nanoparticles in biological analysis such as chemiluminescent analysis based on nucleic acid analysis probes is not investigated

Method used

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  • Luminol direct bonded nano gold nucleic acid analyzing probe and application thereof
  • Luminol direct bonded nano gold nucleic acid analyzing probe and application thereof
  • Luminol direct bonded nano gold nucleic acid analyzing probe and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0083] Taking the method 1) of nucleic acid analysis probe labeling as an example, the preparation method of the nano-gold nucleic acid analysis probe directly bonded to luminol of the present invention comprises the following steps:

[0084] (1) 0.01% (weight) of HAuCl 4 Heat the solution to boiling, under the conditions of condensing and refluxing and fully stirring, add 1.5-2.0mL solution containing 0.01mol / L luminol and 0.01mol / L sodium hydroxide, keep boiling and continue to stir and reflux for 30 minutes , and then remove the heating source and continue to stir for more than 15 minutes to obtain a nano-gold solution with a particle diameter of 14-25nm. The synthesized nano-gold solution was dialyzed at normal temperature for 2 days, and the ultrapure water dialyzate was changed every 8 hours to obtain the nano-gold directly bonded with luminol.

[0085] (2) Streptavidin (Beijing Boaosen Biological Co., Ltd.) was added to the gold nano-sol that had been dialyzed in step ...

Embodiment 4

[0149] Example 4 takes RNA molecules as nucleic acid molecules and target analytes in capture probes as an example to illustrate the specific implementation process of the luminol-directly bonded nano-gold RNA analysis probe and the chemiluminescence analysis method of the present invention.

Embodiment 5

[0150] Embodiment 5 takes the aptamer molecule as the nucleic acid molecule in the capture probe and the corresponding ligand of the aptamer as the target analyte as an example to describe the nano-gold aptamer analysis probe and the aptamer directly bonded to luminol of the present invention The specific implementation process of the chemiluminescence analysis method.

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Abstract

The invention discloses a luminol direct bonded nano gold nucleic acid analyzing probe and a novel chemiluminescence analysis method. The invention provides the luminol direct bonded nano gold nucleic acid analyzing probe, a chemiluminescence analysis method based on the luminol direct bonded nano gold nucleic acid analyzing probe and a kit for constructing the analyzing method; the analyzing probe comprises nucleic acid which is directly bonded by luminol and marked by nano gold; and luminol direct bonded nano gold is obtained by reducing chloroauric acid at one step by luminol. The chemiluminescence analysis method based on the nucleic acid analyzing probe of the invention has the advantages of high sensitivity (for example, specific sequence single-chain DNA detection limit can reach 1.9*10<-16>mol / L), wide linear range, high repeatability, simple operation, low cost, and the like, can be used for the detection of DNA, RNA and adapter corresponding ligands in various samples and has wide application prospect in the fields of clinical diagnosis and treatment, pharmaceutical analysis, food safety detection, environmental monitoring, and the like.

Description

technical field [0001] The invention relates to a nano-gold nucleic acid analysis probe directly bonded with luminol and a method for chemiluminescence analysis using the analysis probe. Background technique [0002] Chemiluminescence analysis is an analytical method to determine the content of the analyte according to the intensity of radiation light generated in the chemical reaction. Because of its high sensitivity, wide linear range, simple instrument, convenient operation, and fast analysis, it has become a very powerful tool in the analysis of metal ions, small molecular organics, amino acids, proteins, and nucleic acids. Among them, the use of chemiluminescence analysis to form analytical probes with nucleic acids and markers to detect specific substances has developed rapidly, and it has become one of the hot spots in the research of chemiluminescence analysis methods in recent years (Li, T.; Li, B.L.; Dong , S.J.Anal.Bioanal.Chem.2007, 389,887; Wang, X.Y.; Zhou, J....

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10G01N21/76
Inventor 崔华柴颖
Owner UNIV OF SCI & TECH OF CHINA
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