Development of fluorescent nanoparticle-labeled lateral flow assay for the detection of nucleic acids

Wang, Y. H. and Nugen, S. R.. (2013) Development of fluorescent nanoparticle-labeled lateral flow assay for the detection of nucleic acids. Biomedical Microdevices, 15 (5). pp. 751-758.

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Abstract

The rapid, specific and sensitive detection of nucleic acids is of utmost importance for the identification of infectious agents, diagnosis and treatment of genetic diseases, and the detection of pathogens related to human health and safety. Here we report the development of a simple and sensitive nucleic acid sequence-based and Ru(bpy)(3)(2+)-doped silica nanoparticle-labeled lateral flow assay which achieves low limit of detection by using fluorescencent nanoparticles. The detection of the synthetic nucleic acid sequences representative of Trypanosoma mRNA, the causative agent for African sleeping sickness, was utilized to demonstrate this assay. The 30 nm spherical Ru(bpy)(3)(2+)-doped silica nanoparticles were prepared in aqueous medium by a novel method recently reported. The nanoparticles were modified by 3-glycidoxypropyl trimeth oxysilane in order to conjugate to amine-capped oligonucleotide reporter probes. The fluorescent intensities of the fluorescent assays were quantified on a mictrotiter plate reader using a custom holder. The experimental results showed that the lateral flow fluorescent assay developed was more sensitive compared with the traditional colloidal gold test strips. The limit of detection for the fluorescent lateral flow assay developed is approximately 0.066 fmols as compared to approximately 15 fmols for the colloidal gold. The limit of detection can further be reduced about one order of magnitude when ``dipstick'' format was used.

Item Type: Article
Additional Information: ISI Document Delivery No.: 211LUTimes Cited: 0Cited Reference Count: 30Wang, Yuhong Nugen, Sam R.USDA 2010-34637-20985; UMass Amherst Center for Hierarchical Manufacturing, a nanoscience shared facility; National Science Foundation CMMI-1025020The authors would like to acknowledge USDA special grant \#2010-34637-20985 for financial support. This project was supported in part by the UMass Amherst Center for Hierarchical Manufacturing, a nanoscience shared facility funded by the National Science Foundation under NSF Grant \#CMMI-1025020.SpringerDordrecht
Uncontrolled Keywords: biosensor
Collections: Nanomanufacturing Research Collection > Nanomanufacturing Nanoscale Science and Engineering Centers > Center for Hierarchical Manufacturing
Depositing User: Robert Stevens
Date Deposited: 27 Mar 2014
Last Modified: 27 Mar 2014 20:21
URI: http://eprints.internano.org/id/eprint/2139

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