A New Fluorescence Immunosensing Method Based on Aptamer-plasmid Complex Amplification
- VernacularTitle:基于核酸适配体-质粒DNA复合物信号放大的荧光免疫传感技术
- Author:
Jing ZHU
;
Yong HUANG
;
Xiaoping JIANG
;
Zhongyang TAN
;
Jianhui JIANG
;
Guoli SHEN
;
Ruqin YU
- Publication Type:Journal Article
- Keywords:
Aptamer;
platelet-derived growth factor;
aptamer-plasmid complex;
immunosense
- From:
Chinese Journal of Analytical Chemistry
2009;37(11):1596-1600
- CountryChina
- Language:Chinese
-
Abstract:
A novel simple,sensitive fluorescence immunosensing method based on aptamer-plasmid complex amplification was developed. This method utilized the specific recognition between antibody and antigen as well as aptamer-plasmid complex and the intercalation of fluorescence dye SYBR Green Ⅰ in the groove of duplex plasmid DNA in detection of Platelet-Derived Growth Factor BB (PDGF-BB). The immunoassay was performed in the microtiter wells in which rabbit anti PDGF-BB antibody was immobilized. The PDGF BB analyte was captured by the primary antibody and then sandwiched by the aptamer-plasmid DNA complex. The introduction of fluorescence dye SYBR Green Ⅰ allows for the detection of the sandwiched immunocomplex of antibody/anigen/aptamer-plasmid complex. Under the optimized conditions of salt concentration,ratio of aptamer to PUC19,and SYBR Green Ⅰ concentration,the proposed method offers a linear detection range from 0.2 μg/L to 200 μg/L with a detection limit of 0.1μg/L.
