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The Research for Nucleic Acid Biosensor Based on Gold Nanoparticles-enzyme Signal Amplilcation System

Author: ZhuQianZuo
Tutor: ZhangYuJing
School: Jilin University
Course: Biochemistry and Molecular Biology
Keywords: Modified gold nanoparticles Enzymatic signal amplification Colorimetric Single-stranded nucleic acid detection
CLC: Q503
Type: Master's thesis
Year: 2011
Downloads: 120
Quote: 0
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Abstract


Gold nanoparticles colorimetric sensors differences in color of gold nanoparticles dispersed state and agglomeration state to design a new type of the intuitive detection technology, the method can be combined with a variety of experimental techniques, testing for a variety of substances. Gold nanoparticles colorimetric method has many advantages: detection is fast and intuitive, low-cost, small capacity and easy to carry. However, such detection methods are also inadequate. Modified gold nanoparticles colorimetric detection of nucleic acids, for example, direct use of this method to detect sensitivity is not very high; Moreover, the current widespread use of biological enzymatic signal amplification technology to improve the sensitivity of the gold nanoparticles colorimetric method, but these methods is extremely big limitations, can not meet all of the nucleic acid detection system. Therefore, the design of a sensitive, for any nucleic acid sequence, operating a simple detection system is an important research subject of the present. The same enzyme signal amplification technology with modified gold nanoparticles colorimetric combination to detect the target nucleic acid, two have been devised for any single stranded nucleic acid detection methods. The first one is the alternative amplification chain technology is introduced into the gold nanoparticles colorimetric methods: triggered by the addition of the target nucleic acid sequence strand displacement amplification reaction carried the product hybridized nucleic acid can be complementary to the connection chain Linker, block modified gold nanoparticles agglomeration ; In contrast, when the target nucleic acid, linker cause modified gold nanoparticle agglomeration discoloration. This method has the advantages of intuitive, simple, low cost, without any background, a detection limit of 1 pM. Another method is the use of the restriction endonuclease cycle digested amplified signal: The system needs to HincII enzyme, a a phosphorothioate hairpin probe, the connection chain Linker as well as the nucleic acid modified gold nanoparticles. Hairpin probe and the target nucleic acid interactions change their idea to open the \nucleic acid is present, the hairpin probe is very stable and can not lead to a restriction cycle reaction, modified gold nanoparticle agglomeration discoloration. Detecting target nucleic acid of the design concept is very simple and sensitive method specificity, limit of detection was 5 pM, compared to the the stage direct use nucleic acid modification of the nano gold \The differences due to the normal target nucleic acid and the mutated nucleic acid and hairpin probe hybridization Tm value, using this principle, the experimental method is also applicable to the detection of nucleic acid base mutation. This paper aims to design an intuitive sensitivity for any single-stranded nucleic acid detection methods, are decorated with gold nanoparticles colorimetric detection of nucleic acid promote the application of the method to lay a foundation for this simulation test. In addition, these new methods can be easily with the current conventional analytical techniques combined, the sensing area can be expanded to include a protein, a nucleic acid aptamer of small molecules, such as metal ions.

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