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作 者:王艳君[1] 李永生[2] 杨全玉[1] 黄燚[1] 唐静[1] 高秀峰[1]
机构地区:[1]四川大学华西基础医学与法医学院,四川成都610041 [2]四川大学化学工程学院,四川成都610065
出 处:《光谱学与光谱分析》2009年第1期165-168,共4页Spectroscopy and Spectral Analysis
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(0082204127092)资助
摘 要: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.
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