Title |
Novel Competitive Chemiluminescence DNA Assay Based on Fe3O4@SiO2@Au-Functionalized Magnetic Nanoparticles for Sensitive Detection of p53 Tumor Suppressor Gene
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Published in |
Applied Biochemistry and Biotechnology, June 2018
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DOI | 10.1007/s12010-018-2808-1 |
Pubmed ID | |
Authors |
Linyu Wang, Manwen Yao, Xiangyi Fang, Xi Yao |
Abstract |
A simple, rapid response time and ultrahigh sensitive chemiluminescence (CL) DNA assay based on Fe3O4@SiO2@Au-functionalized magnetic nanoparticles (Au-MNPs) was developed for detection of p53 tumor suppressor gene. In this study, 2',6'-dimethylcarbonylphenyl-10-sulfopropyl acridinium-9-carboxylate 4'-NHS ester (NSP-DMAE-NHS), as a new kind of highly efficient luminescence reagent, was immobilized on the complementary sequence of the wild-type p53 (ssDNA) to improve the detection sensitivity. The optimal concentration of ssDNA-(NSP-DMAE-NHS) conjugates mixed with the wild-type p53 (wtp53) samples respectively. Then, the wtp53-Au-MNPs conjugates were added to continue the competitive reaction in the above solution. Subsequently, the Au-MNPs separated under magnetic field, measured by a homemade luminescent measurement system. Under optimal conditions, the method exhibited ultrasensitive sensitivity with a detection limit of 0.001 ng mL-1 (0.16 pM), a wide range of liner response from 0.001 ng mL-1~6.6 μg mL-1. Therefore, the immunomagnetic nanocomposites-based detection strategy was rapid, low-cost, and highly sensitive that can be easily extended to the early diagnosis of cancer development and monitoring of patient therapy. |
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