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Study on the Detection of Avian Influenza Virus Genotype Based on PAMAM Modified Electrode

Author: ZhuXiangBin
Tutor: AiShiYun
School: Shandong Agricultural University
Course: Environmental Science
Keywords: Biosensor Carbon nanotubes Dendrimer DNA Guanine Avian influenza virus
CLC: S854.43
Type: Master's thesis
Year: 2010
Downloads: 34
Quote: 1
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Abstract


DNA electrochemical biosensor is a means of electrochemical biosensor analysis and detection of DNA structure, with a high degree of molecular recognition accuracy, high sensitivity, ease of integration and automation, etc., especially suitable for the surface process modified electrode The electron transfer process has been widely applied in the field of genetic diagnosis, infectious disease detection, gene expression, forensic and environmental monitoring. In the the sensor build process, how the effective use of biological molecules fixed technology and choose the fixed material to determine DNA electrochemical biosensor stability, selectivity and sensitivity of the main performance. This thesis explores design three novel electrochemical biosensors based on guanine the G oxidation signal the change to the initial detection of the avian influenza virus DNA immobilization and hybridization method, using cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy to characterize them, and DNA electrochemical biosensor for the detection of avian influenza virus. The study mainly divided into the following three parts: (1) the use of cobalt hexacyanoferrate (CoHCF-PAMAM) - dendrimer composite modified glassy carbon electrode (GCE), preparation of the label-free detection of gene mutations in a novel DNA electrochemical sensor . Sensor dendrimer layer significantly increased the amount of single-stranded DNA probe fixed, cobalt hexacyanoferrate layer increases the oxidation of guanine signal, that the sensor can be sensitive to identify single nucleotide mismatch in gene sequence, having good selectivity and sensitivity. 7.6 × 10 -11 to 3.05 × 10 -8 mol / L concentration range, guanine (G), the oxidation peak current difference between the concentration of the mutant gene was a good linear relationship, a detection limit of 1.0 × 10 -11 mol / L (66 pg / mL) (S / N = 3). (2) the use of lead dioxide (PbO 2 ), multi-walled carbon nanotubes (MWNTs) and hydrophobic room temperature ionic liquid (RTIL) 1 - butyl - 3 - methylimidazolium hexafluorophosphate (BMIMPF6) composite modified glassy carbon electrode the (PbO 2 -MWNT-RTIL/GCE), prepared by a novel electrochemical sensors used simultaneously or separately detect guanine (G) and adenine (A). When the modified electrode after to enriched 120 s under the applied voltage 0.30 V at pH 7.0 phosphate buffer solution, differential pulse voltammetry (DPV) detection, guanine and adenine, respectively, 0.60 V and 0.89 V at oxidation peak. Compared with the bare electrode, PbO 2 -MWNT-RTIL/GCE not only significantly improve the electrochemical oxidation of guanine and adenine peak current, and reduce the oxidation peak potential, suggesting that PbO 2 , the MWNT and RTIL synergies can be good to improve the sensitivity of detection of guanine and adenine. The experimental results show that the PbO 2 -MWNT-RTIL/GCE has good stability, excellent electrocatalytic properties and enrichment effect, and can simultaneously detect guanine and adenine (oxidation peak potential difference of 0.29 V). The modified electrode under the best conditions, guanine and adenine detection limit of 1.0 ng / mL and 4.0 ng / mL (S / N = 3, signal-to-noise ratio). Guanine and adenine herring sperm DNA detectable by this method achieved satisfactory results. (3) Preparation of the use of a label-free method to detect the DNA sequence of the avian influenza virus, a novel DNA electrochemical biosensor. First of all, the multi-walled carbon nanotubes and cobalt phthalocyanine nano-composites (MWNTs-CoPc) and 4.0G dendrimer (PAMAM G4.0) modified glassy carbon electrode, then G4 PAMAM bridge DNA probe fixed to the modified electrode. Observed by differential pulse voltammetry the guanine oxidation change in the signal to detect the hybridization reaction, to avoid the use of external indicator. Under the optimum conditions, the number of pairs of the electrode obtained in the range of 0.01 to 500 ng / mL, modified guanine oxidation signal difference (ΔI p ) with the complementary target gene concentration complementary target gene concentration a linear relationship, the correlation coefficient was 0.998, and the detection limit of 1.0 pg / mL.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary clinical medicine > Veterinary diagnostics > Laboratory diagnostic method
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