dna电化学生物传感器检测乳腺癌转移标志基因的实验分析word格式论文.docx

Experimental studies on the DNA electrochemical biosensor for detection of marker genes in breast cancer metastasisAbstractObjective:To provide the scientific basis for detection of breast cancer marker genes, a electrochemical sensor was prepared and investigated, and the performance of the sensor also was studied..To establish a DNA electrochemical biosensor for detection of mammaglobin(MG) in breast cancer metastasis.To establish a DNA electrochemical biosensor for detection of cytokeratin 19 (CK19) in breast cancer metastasis, and to achieve early diagnosis of breast cancer metastasis.Methods:Polybromothymol blue (BTB)-modified electrode was prepared using cyclic voltammetry in pH 5.0 PBS containing 0.2 mM BTB in the potential window of -0.2 to 1.6V at a scan rate of 100 mVs-1 for 15 cycles. The electrochemically synthesized film was investigated using electrochemical impedance spectroscopy. Using ascorbic acid, dopamine and uric acid as the model of electrical activity, the performance of the sensor was studied by simultaneous determination of these three compounds in phosphate buffer solution (pH 5.0).Using glassy carbon electrode as the substrate electrode, we designed a length of 25 bp single-stranded DNA fragment of mammaglobin as probe, and then we immobilized the probe on the electrode surface by covalent Bonding. After this, we detected the nature of electrode by electrochemical impedance technique before and after DNA hybridization, and optimized the experimental conditions, constructed DNA electrochemical biosensor based on the new method of detecting breast gene metastasis.we used the sener to detect the actual PCR amplification products in clinical breast cancer patient, and we also compared the fact with conventional gelelectrophoresis.Using gold electrode as the substrate electrode, we designed a length of 25 bp single-stranded DNA fragment of cytokeratin 19 as probe,and then we immobilized the probe on the electrode surface by self-assembly metho