A versatile and sensitive colorimetric assay that allows the rapid detection of small-molecule targets using the naked eye is demonstrated. The working principle of the assay integrates aptamer-target recognition and the aptamer-controlled growth of gold nanoparticles (Au NPs). Aptamer-target interactions modulate the amount of aptamer strands adsorbed on the surface of aptamer-functionalized Au NPs via desorption of the aptamer strands when target molecules bind with the aptamer. Depending on the resulting aptamer coverage, Au NPs grow into morphologically varied nanostructures, which give rise to different colored solutions. Au NPs with low aptamer coverage grow into spherical NPs, which produce red-colored solutions, whereas Au NPs with high aptamer coverage grow into branched NPs, which produce blue-colored solutions. We achieved visible colorimetric response and nanomolar detection limits for the detection of ochratoxin A (1 nM) in red wine samples, as well as cocaine (1 nM) and 17β-estradiol (0.2 nM) in spiked synthetic urine and saliva, respectively. The detection limits were well within clinically and physiologically relevant ranges, and below the maximum food safety limits. The assay is highly sensitive, specific, and able to detect an array of analytes rapidly without requiring sophisticated equipment, making it relevant for many applications, such as high-throughput drug and clinical screening, food sampling, and diagnostics. Furthermore, the assay is easily adapted as a chip-based platform for rapid and portable target detection.
In this article, we report on direct detection of microRNAs (miRNAs) on a microarray by differential interference contrast (DIC) imaging technique. While the best resolution achieved with a fluorescence scanner is ∼1 μm, the DIC imaging technique adopted in our study offers the possibility of imaging individual reporting gold nanoparticles, or, in other words, individual miRNA strands. Due to its unrivalled resolution, the present technique could detect as low as 300 copies of target miRNAs in a sample volume of 1.0 μl. With the greatly improved sensitivity, the amount of total RNA needed in the assay is reduced to only a few nanograms, offering an excellent opportunity for fast and direct miRNA profiling without engaging any labeling and amplification procedure. Expression patterns of hsa-let-7 family members in healthy versus cancer cells analyzed on our microarray, are found to be consistent with the patterns obtained on a commercial microarray and those reported in the literature.
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