Goldview标记的DNA荧光毛细生物传感器的研究

Research on DNA Fluorescence Capillary Biosensor Marked by Goldview

对Goldview（GV）作为荧光标记物的DNA荧光毛细生物传感器进行了研究。以荧光毛细分析法（fluorescence capillary analysis,FCA）为基础,在毛细管内壁通过Poly-l-lysine将20-mer-ssDNA探针固定,制成DNA荧光毛细生物传感器（DNA fluorescence capillary biosensor,DNA-FCB）,DNA-FCB与互补靶DNA杂交,通过GV染色后,检测杂交产物的荧光强度,实现对靶DNA的定性和定量分析。样品用量12μL,靶DNA的浓度在0.4-4μmol·L^-1（2.4-24 mg·L^-1）范围内和荧光强度有良好的线性关系（y=65.911x＋3.994 4,r=0.998 9）;RSD〈3.5%,检出限0.39μmol·L^-1（2.2 mg·L^-1）,能达到定量检测靶DNA的目的。用DNA-FCB测定靶DNA操作简便,试样、试剂用量少,测定成本极低,能大大减少环境污染。

Goldview marked DNA fluorescence capillary biosensor was studied in the present paper.Based on fluorescence capillary analysis （FCA）,the DNA biosensor uses capillary as immobilization carrier and detection carrier of DNA probe.Probes（20-mer-ssDNA） were immobilized on the inner wall of capillary by poly-l-lysine,and DNA fluorescence capillary biosensor（DNA-FCB） was made.After being hybridized with complementary target DNA and dyed by Goldview,the target DNA was qualified or quantified by detecting the fluorescence density of the Goldview using F-4500 spectrofluorometer.The sample volume was 12 μL.The concentration of the target DNA showed good linearity with the fluorescence intensity in the range of 0.4-4 μmol·L^-1（2.4-24 mg·^L-1）（y=65.911x＋3.994 4,r=0.998 9）.The RSD was lower than 3.5%.The concentration detection limit of the target DNA was 0.39 μmol·L^-1（2.2 mg·L^-1）.The DNA-FCB can be used to qualify or quantify the target DNA.It＇s advantages are simplicity of manipulation,thimbleful of sample and reagent volumes,repeated use of capillary,and the lowest test cost.By using DNA-FCB to qualify the target DNA,we can consumedly decrease the pollution of the environment.