肝门部胆管癌术前MDCT诊断及可切除性分析临床应用研究-普通外科学专业论文.docxVIP

广州医学院硕士学位论文刘杰文 广州医学院硕士学位论文 刘杰文 PAGE PAGE 7 To establish more stabilized nanogold-labeled microarray technology，we optimized the spotting buffer，the concentration of nanogold and the time of silver enhancement. In the meantime，we studied the effect of pre-hybridization on hybridization signal，as well as the detection limit of the nanogold-labeled microarray.Application of the labelled Nanogold microarray to detect 11 kinds of genes for prostate cance targets (IGF1, P27, AR, PSMA, KLK3, PCNA, Ki-67, KLK1, KLK2, TMPRSS2, SREBF2) and clinical specimens of prostate cancer, study on each of the gene expressions. RESULTS: The results showed that Labelled Nanogold Microarray detected when the sensitivity of the single-stranded DNA reached to 80 fmol/L ,the hybridization had a good specificity. Among the three kinds of spotting buffer，50%DMSO was the best. Pre-hybridization led to 70% loss of hybridization signal. The time of silver enhancement we chose was 15 minutes，at which the target signal was clear，while the background was low. Under the condition , Application of the labelled Nanogold microarray to detect 11 kinds of genes for prostate cance targets (IGF1, P27, AR, PSMA, KLK3, PCNA, Ki-67, KLK1, KLK2, TMPRSS2, SREBF2) and clinical specimens of prostate cancer, each upregulation gene probe point presented the obvious chromogenic expression，and the different shades.Downregulation gene and the negative control presented no chromogenic expression, This can be figure out by human eyes,to achieve qualitative and quantitative results. CONCLUSION: 1、Under the condition , the best kinds of spotting buffer was 50%DMSO, Pre-hybridization led to 70% loss of hybridization signal. The time of silver enhancement we chose was 15 minutes，at which the target signal was clear， while the background was low,the sensitivity of the single-stranded DNA reached to 80 fmol/L.Can be used as a relatively reliable detection of gene chip platform,with high sensitivity and good specificity ,low