【作者】 夏骊； 张利娟； 宋重阳； 庞代文； 唐宏武；

【Author】 Li Xia;Li-Juan Zhang;Chong-Yang Song;Dai-Wen Pang;Hong-Wu Tang;Key Laboratory of Analytical Chemistry for Biology and Medicine(MOE),College of Chemistry and Molecular Sciences,Wuhan University;

【机构】 武汉大学化学与分子科学学院生物医学分析化学教育部重点实验室；

【摘要】 酶联免疫吸附试验（Enzyme-linked immunosorbent assay,ELISA）是一种利用酶联抗体结合抗原的特异性检测吸附在固相载体表面抗原的方法,因其在蛋白质测定中具有快速、灵敏、简便等优点而受到广泛应用^[1,2]。我们将该方法应用于DNA检测,实现了DNA含量的可视化分析。首先将核酸探针连接在96孔酶标板表面,然后用其捕获生物素化的互补DNA,最后加入辣根过氧化物酶标记的链霉亲和素,通过其催化底物显色对待测核酸进行定量分析。此方法在DNA可视化测定中具有操作简便、灵敏度高等优点,检测限低至60 fM。更多还原

【Abstract】 The need for analysis of target nucleic acids has driven the development of membrane hybridizations.This method is time-consuming considering that it requires long incubation and washing time to allow reagents to permeate into the membrane.Here,we present a much more rapid and simpler technique applied to nucleic acid hybridization assays in a format synonymous with common enzyme-linked immunosorbent assays(ELISA).Nucleic acid molecules are coupled to a 96 well microplate surface and are used to capture biotinylated target DNA molecules.The wells are subsequently incubated with a streptavidin-enzyme conjugate followed by the addition of the substrate.This labeled nucleic acid could be detected based on a positive colorimetric assay with detection limit 60 fM.更多还原