title: Development of highly fluorescent silica nanoparticles chemically doped with organic dye for sensitive DNA microarray detection.
abstract: Increasing the sensitivity in DNA microarray hybridization can significantly enhance the capability of microarray technology for a wide range of research and clinical diagnostic applications, especially for those with limited sample biomass. To address this issue, using reverse microemulsion method and surface chemistry, a novel class of homogenous, photostable, highly fluorescent streptavidin-functionalized silica nanoparticles was developed, in which Alexa Fluor 647 (AF647) molecules were covalently embedded. The coating of bovine serum albumin on the resultant fluorescent particles can greatly eliminate nonspecific background signal interference. The thus-synthesized fluorescent nanoparticles can specifically recognize biotin-labeled target DNA hybridized to the microarray via streptavidin-biotin interaction. The response of this DNA microarray technology exhibited a linear range within 0.2 to 10 pM complementary DNA and limit of detection of 0.1 pM, enhancing microarray hybridization sensitivity over tenfold. This promising technology may be potentially applied to other binding events such as specific interactions between proteins.
authors.affiliation: Institute for Environmental Genomics, Department of Botany and Microbiology, University of Oklahoma, Norman, OK 73019, USA. liuah@qibebt.ac.cn
authors.forename: Aihua
authors.initials: A
authors.lastname: Liu
authors.forename: Liyou
authors.initials: L
authors.lastname: Wu
authors.forename: Zhili
authors.initials: Z
authors.lastname: He
authors.forename: Jizhong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001710:Biotin / Q000378:metabolism; D002417:Cattle; D004247:DNA / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D012710:Serum Albumin, Bovine; D001710:Biotin; D012822:Silicon Dioxide; D004247:DNA; D019809:Streptavidin