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Microchip Capillary Electrochemical with Electrochemical Detection for Tumor Markers and DNA

Author: SuMingMing
Tutor: ZhangXiaoRu
School: Qingdao University of Science and Technology
Course: Physical and chemical
Keywords: Microchip Capillary Electrophoresis Electrochemical Detection Horseradish peroxidase Tumor markers DNA
CLC: R446
Type: Master's thesis
Year: 2010
Downloads: 37
Quote: 0
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Abstract


This work was to establish a microchip capillary electrophoresis with electrochemical detection of tumor markers and DNA methods. By o-aminophenol - hydrogen peroxide - horseradish peroxidase (OAP-H 2 O 2 -HRP) system, the use of microchip capillary electrophoresis separation with electrochemical detection techniques detection of tumor markers and DNA. The first chapter is an introduction, a brief introduction of the microchip capillary electrophoresis and analysis on the current commonly used in microfluidic electrochemical detection methods are briefly reviewed. Introduced capillary electrophoresis chip development and research status quo. The second chapter describes the PDMS / glass composite chip and the electrode design and processing. The composite chip the glass substrate and a PDMS chip. This section briefly describes processing and PDMS chip electrodes on the glass substrate design and processing. Chapter III chip capillary electrophoresis o-aminophenol (OAP) - H 2 O 2 - horseradish peroxidase (HRP) enzyme immunoassay system for a more detailed study, the optimal reaction conditions, the determination of free HRP linear range of 4.5 × 10 -9 7.8 × 10 -13 mol / L, the detection limit was 1.6 × 10 -13 mol / L (S / N = 3). Chapter IV chip capillary electrophoresis using the o-aminophenol (OAP) - H 2 O 2 - horseradish peroxidase (HRP) enzyme immunoassay with electrochemical detection system the alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG). This method has been successfully used for alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG) is detected. AFP and HCG linear ranges were 1.5 6 6.6 ng / mL and 1.7 1 32.5 ng / mL, detection limits were 0.48 ng / mL and 0.30 ng / mL. Chapter V-chip capillary electrophoresis using the o-aminophenol (OAP) - H 2 O 2 - horseradish peroxidase (HRP) enzyme immunoassay with electrochemical detection system of 27 and 39 nucleotides of the target DNA. This method uses biotinylated DNA to a modified pro-horseradish peroxidase nuclide (avidin-HRP) to form HRP-labeled DNA probe, followed by DNA strand and its complementary hybridization. After the hybridization reaction, the mixture including the HRP-labeled double-stranded DNA (dsDNA-HRP), an excess of HRP-labeled single-stranded DNA (ssDNA-HRP) and the remaining pro-nuclides modified horseradish peroxidase (avidin-HRP), Finally, the chip capillary electrophoresis for separation. This method is successful separation and detection of 27, 39 bases of the target DNA, their linear range was 3.0 × 10 -11 -1.8 × 10 -9 mol / L and 4.0 × 10 -11 -1.4 × 10 -9 mol / L, the detection limit was 8 × 10 -12 mol / L and 1.2 × 10 -11 mol / L; and large field coli genomic DNA for the separation and detection of PCR products.

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