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目的:建立简便快速、稳定的提取胰腺癌患者外周血循环DNA的固相载体法。方法:异硫氰酸胍-硅胶法提取DNA,并利用灵敏度极高的新型低毒荧光染料SYBR GreenⅠ对所提取的DNA进行显示。结果:异硫氰酸胍-硅胶法可用于提取血清DNA,且重复性好,SYBR GreenⅠ染色显示胰腺癌患者外周血循环DNA片段大小主要包括完整的基因组DNA、2000bp左右片断或两者兼而有之(而正常者几乎没有)。结论:胰腺癌患者外周血中存在较高水平的循环DNA,异硫氰酸胍-硅胶法提取及SYBR GreenⅠ染色显示外周血循环DNA简便实用,具有较大的临床开发价值。
Objective: To establish a simple, rapid and stable solid phase extraction method for DNA extraction from peripheral blood of patients with pancreatic cancer. Methods: The DNA was extracted by guanidine isothiocyanate - silica gel method and the extracted DNA was displayed by SYBR Green Ⅰ, a new type of low toxicity fluorescent dye with high sensitivity. Results: Guanidine isothiocyanate - silica gel method can be used to extract serum DNA with good repeatability. The SYBR Green Ⅰ staining showed that the size of circulating DNA fragments in peripheral blood of patients with pancreatic cancer mainly consisted of complete genomic DNA, fragments of about 2000bp or both (And normal almost no). Conclusion: There is a high level of circulating DNA in the peripheral blood of patients with pancreatic cancer. Extraction with guanidinium isothiocyanate - silica gel and SYBR Green Ⅰ staining showed that DNA in peripheral blood is simple and practical, which has great value in clinical development.