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Method of colorimetric detection of target DNA by combining nanometer gold with polythiophene ramification

A technology of polythiophene derivatives and detection methods, which is applied in the field of target gene detection, can solve problems such as high cost, and achieve the effects of simple operation and convenient detection

Inactive Publication Date: 2010-09-01
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is usually necessary to modify oligonucleotide probes on the surface of gold nanoparticles to achieve the purpose of specific recognition of target biomolecules, so the cost is relatively expensive.

Method used

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  • Method of colorimetric detection of target DNA by combining nanometer gold with polythiophene ramification
  • Method of colorimetric detection of target DNA by combining nanometer gold with polythiophene ramification
  • Method of colorimetric detection of target DNA by combining nanometer gold with polythiophene ramification

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Experimental program
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Effect test

Embodiment 1

[0043] (1) Preparation of detection reagents and probes

[0044] (1) Preparation of nano-gold solution

[0045] Prepare a solution with a concentration of 1 mM HAuCl4 and a solution with a concentration of 38.8 mM, trisodium citrate. Heat the HAuCl4 solution to 130°C, ensure sufficient stirring during the heating process, quickly add 25ml of the newly prepared trisodium citrate solution (pay attention to heat preservation during this process, do not let the HAuCl4 solution cool down) after about 20 minutes, and continue stirring until the solution cools down At room temperature, a wine-red solution is formed. Filter the solution with a 0.22 μm nitrocellulose nylon membrane to obtain a 13nm nano-gold solution with uniform particles, and store it at 4°C for later use.

[0046] (2) Design and synthesis of H1N1 DNA detection probes

[0047] The invention uses the H1N1 (GenBank: CY048965.1) sequence as a target gene template to establish an analysis method for direct gene detect...

Embodiment 2

[0062] Comparison of the method for detecting target DNA in combination with nano-gold combined with polythiophene derivatives provided by the present invention and nano-gold and polythiophene derivatives alone

[0063] 1. Colorimetric detection of single-stranded and double-stranded DNA based on nano-gold solution

[0064] 1.1 Preparation of single-stranded, double-stranded and single-base mutant DNA solutions

[0065] Take an Ependoff tube and add 3 μL of deionized water, 1 μL of H1N1 DNA probe, 1 μL of non-complementary DNA (HCV DNA control) and 5 μL of 0.2M NaCl / 10mM PB solution, mix well, and prepare non-double-stranded DNA solution (single-stranded DNA) Take another tube and add 1 μL of deionized water, 2 μL of H1N1 DNA probe, 2 μL of complementary target H1N1 DNA, and 5 μL of 0.2M NaCl / 10mMPB solution to prepare a double-stranded DNA solution with a concentration of 20 μM.

[0066] 1.2. Detection of complementary target DNA

[0067] At room temperature, take 5 μL each...

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Abstract

The invention relates to a method of the colorimetric detection of target DNA by combining nanometer gold with polythiophene ramification, which comprises the following steps of: preparing a detection reagent (the nanometer gold and the polythiophene ramification) and a probe; and detecting the sequence of the target, etc. Based on the principle that a signal cascade is amplified due to the change of the color since the stability of the NDA-nanometer golden solution is changed when the polythiophene ramification is combined with the ssDNA / dDNA, the method realizes the high-sensitivity and fast detection of the DNA under the double action of a color converting developer and an amplified label, thereby building an analysis platform which directly detects the gene by combining the nanometer gold with the polythiophene ramification. The method has simple operation, does not need special instrument and equipment, has the characteristics of specificity, speediness and high sensitivity, and can be applied to the diagnosis and the detection of the target DNA in the clinical laboratory medicine and the environment.

Description

technical field [0001] The invention relates to a method for colorimetric detection of target DNA in combination with nano gold and polythiophene derivatives. More precisely, it particularly relates to a method for sensitive and rapid detection of target DNA by composite gold nanoparticles and polythiophene. It belongs to the field of target gene detection. Background technique [0002] Human, viral and bacterial specific nucleic acid sequence detection is becoming increasingly important in disease diagnosis. Especially in recent years, outbreaks caused by some pathogens have brought serious losses to human health and social economy. Therefore, the rapid and accurate diagnosis of pathogens is not only of great significance to the prevention and control of infectious diseases, but also plays a positive role in the guidance of clinical diseases. Many techniques, including electrochemical, fluorescent, electrophoretic, electrochemiluminescent, enzymatic, surface plasmon reso...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/52G01N21/33
Inventor 毛红菊王树郭青川娄新徽张宏莲金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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