全文下载：2012010042
牛淑妍， 娄晓飞， 渠丽景
（青岛科技大学 化学与分子工程学院，山东 青岛 266042）
摘要： 利用纳米金放大的方法，将修饰荧光团的信号DNA连接在纳米金表面，纳米金通过HBV-DNA与磁珠连接，1,4-二硫苏糖醇（DTT）可以替代信号DNA连接在纳米金表面，将荧光素标记的信号DNA释放于溶液中，测定溶液的荧光强度可以得到HBV-DNA的浓度。通过条件优化实验确定了最佳实验条件：生物素与亲和素最佳结合时间为25 min，DNA的最佳杂交时间为15 min。测定HBV-DNA的线性范围为3.0 × 10-13～1.2 × 10-12 mol·L-1，线性相关系数r= 0.991 4，检测限为2.18 × 10-14 mol·L-1（S/N=3）。
关键词： HBV-DNA； 纳米金； 荧光光谱
中图分类号： O 657. 32文献标志码： A
Gold Nanoparticles Amplification Assay for Fluorescence-detection of Hepatitis B Virus DNA
NIU Shu-yan， LOU Xiao-fei， QU Li-jing
(CollegeofChemistryand Molecular Engineering,QingdaoUniversityof Science and Technology,Qingdao266042,China)
Abstract: Using a highly ultrasensitive fluorescence-detection method based on gold nanoparticles and magnetic beads assisted by dithiothreitol (DTT) could detect the target DNA. The detection of HBV-DNA was achieved by monitoring fluorescence signals obtained from the adsorbed thiolated oligonucleotides modified with fluorophore which were liberated from the gold nanoparticle surface. The best reaction conditions were obtained through the study on the effect of different reaction time on the system’s fluorescent intensity. The optimal reaction time of biotin and streptavidin was 25 min. 15 min was chosen as the hybridization time of capture probes and target DNA. In the system of fluorescence spectroscopy based on gold nanoparticles, the HBV-DNA could be quantified in the range of 3.0 × 10-13—1.2 × 10-12 mol·L-1 and the detection limit was 2.18 × 10-14 mol·L-1（S/N=3）.
Key words: HBV-DNA; gold nanoparticles; fluorescence spectroscopy
收稿日期：2011-04-12
基金项目： 国家自然科学基金项目(21075073)；山东省自然科学基金重点项目(ZR2010BZ006).
作者简介： 牛淑妍 (1963—)，女，教授.
