Cy5标记的DNA荧光毛细生物传感器的研究  

Study on DNA Fluorescence Capillary Biosensor Labeled with Cy5

作　　者：王艳君[1] 李永生[2] 高秀峰[2] 杨全玉[2] 

机构地区：[1]四川省卫生学校,四川成都610100 [2]四川大学,四川成都610041

出　　处：《标记免疫分析与临床》2011年第3期182-185,共4页Labeled Immunoassays and Clinical Medicine

基　　金：教育部留学回国人员科研启动基金项目(2300448);四川大学214振兴计划科研启动基金(N0.0082204127092)

摘　　要：探索了以Cy5（吲哚-5-菁）作为荧光标记物的DNA荧光毛细生物传感器的可行性。以荧光毛细分析法（fluorescencecapillary analysis,FCA）为基础,在毛细管内壁通过poly-l-lysine将20-mer-ss DNA探针固定,制成DNA荧光毛细生物传感器（DNA fluorescence capillary biosensor,DNA-FCB）,DNA-FCB吸入含Cy5标记的靶DNA液杂交,通过检测杂交产物的荧光强度,实现对靶DNA的定性和定量分析。结果显示,样品用量5μL,Cy5标记的靶DNA浓度在0.2~1.2μmol/L（2.4~24mg/L）范围内和荧光强度有良好的线性关系（Y=129.61X＋21.233,r=0.9970）;RSD〈3.5%;检出限为0.18μmol/L。结论：以Cy5作为荧光标记物的DNA荧光毛细生物传感器能达到定量检测靶DNA的目的。该方法操作简便,试样、试剂用量少,毛细管及DNA-FCB可循环使用,测定成本极低,能大大减少环境污染。To investigate the feasibility of DNA fluorescence capillary biosensor labeled with Cy5,based on fluorescence capillary analysis（FCA）,the DNA biosensor using capillary as immobilization and detection carrier of DNA probe（20-mer-ssDNA） were immobilized on the inner wall of capillary by poly-l-lysine to make DNA fluorescence capillary biosensor（DNA-FCB）.The DNA-FCB was hybridized with complementary target DNA labeled by Cy5.The target DNA was qualified or quantified by detecting the fluorescent density of the Cy5 using F-4500 spectrofluorometer.The results showed that the concentration of the target DNA had good linearity with the fluorescent intensity in range of 0.2～1.2μmol/L when sample volume was 5μL.RSD was lower than 3.5%.The concentration detection limit of the target DNA was 0.39μmol/L.The DNA-FCB can be used to qualify or quantify the target DNA.DNA-FCB is simple with advantages of lower sample and reagent volumes,repeated use of capillary,the lowest test cost,and reduce the pollution to environment.

关 键 词：DNA 荧光毛细生物传感器 Cy5 DNA-FCB 

分 类 号：Q503[生物学—生物化学]