Biofunctional organic nanocrystals for quantitative detection of pathogen deoxyribonucleic acid |
| |
Authors: | Chan Cangel Pui-yee Tzang Lawrence Chi-hung Sin King-keung Ji Sheng-lin Cheung Kwan-yee Tam Tze-kin Yang Michael Meng-su Renneberg Reinhard Seydack Matthias |
| |
Institution: | a Department of Chemistry, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, SAR, Hong Kong b Department of Biology and Chemistry, The City University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong c Division of Bioengineering and Department of Chemical & Biomolecular Engineering, National University of Singapore, Singapore d 8sens.biognostic GmbH, Robert-Roessle-Strasse 10, D-13125 Berlin, Germany |
| |
Abstract: | Advances in nanotechnology have had significant impacts in the field of biodiagnostics. In this study, we describe the novel application of dissolvable, organic and biofunctional nanocrystals for the quantitative detection of a PCR product. Fluorescein diacetate (FDA), a fluorogenic precursor of fluorescein, was milled in a solution of a polymeric surfactant to create a stable, nanosized colloid with an interface for coupling streptavidin molecules. The application of these particulate labels for the quantitative detection of biotinylated human papillomavirus (HPV) DNA, amplified in a standard PCR procedure, was demonstrated. After the affinity reaction, the FDA molecules were dissolved and concomitantly converted into fluorescein. This approach resulted in a high selectivity, short incubation times and a sensitivity up to 147 times greater than obtained from state-of-the-art, directly fluorescent-labeled streptavidins. This innovative method offers rapid detection of small amounts of nucleic acids because less target material and thus fewer PCR cycles are required. |
| |
Keywords: | Fluorescein diacetate Fluorescein 5-isothiocyanate Horseradish peroxidase Human papillomavirus |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|