ReviewGold nanoparticle probes for the detection of nucleic acid targets☆
Introduction
Gold nanoparticles have been used in biotechnology over the last 4 decades as immunocytochemical probes as well as biological tags [1], [2]. During the last decade, however, there has been an increasing interest in using nanoparticles in DNA detection [3]. Recent advances in functionalizing particles with oligonucleotides and tailoring their surface properties have paved the way for the development of a series of new and practical biodetection systems [4], [5]. Combining new capabilities for controlling particle size and composition with versatile surface modification approaches allows for the design of optically and chemically encoded nanoparticle probes. These probes be used in biomolecule detection assays with numerous advantages when compared to conventional assays [6], [7]. In addition to the optical and recognition properties imparted by surface molecules, metal nanoparticles also conduct electricity and are quite useful as catalysts due to their high surface area. For nucleic acid targets, all of these properties have allowed for the design of numerous assays. Examples are reviewed here and compared to the most relevant conventional techniques, where fluorescent and radioactive detection readout methods are the most common (e.g., PCR, RT-PCR, northern blot, southern blot, high density microarrays). Due to space limitations, readers are referred to the literature for reviews of other nanoparticle detection methods [2], [3], [6], [7].
Section snippets
Homogeneous detection
A turning point in the bio-analytical applications of gold nanoparticles in DNA sensing was the discovery in 1996 that gold nanoparticles (Au-NPs) could be surface-functionalized with thiolated oligonucleotides (DNA-Au-NPs) and stabilized in aqueous biological buffers [4]. This discovery allowed researchers to tailor the properties of the nanoparticle probes, as well as identify new properties derived from either the probe itself or the dense loading of oligonucleotides on the particle surface.
Heterogeneous detection
In an effort to develop systems with higher sensitivities, there was a shift of interest to the development of assays that support highly multiplexed surface-based readouts. The power of microarray technology is the ability to detect hundreds to thousands of DNA or RNA sequences in a single assay and quantitatively compare two samples [14], [15]. As a first step, the scanometric assay was developed, which is a sandwich assay involving an immobilized capture strand on a chip, a target sequence,
The bio-barcode assay: homogeneous meets heterogeneous
The polymerase chain reaction (PCR), when set up properly, is powerful enough to amplify a target nucleic acid sequence from even a single cell, such that large quantities are available for downstream applications, sequence confirmation, and quantification. Real-time PCR (RT-PCR) has become popular, which allows for sequence-specific comparative quantification of DNA expression and SNP analysis [30]. For nucleic acids, PCR is powerful with respect to sensitivity but carries with it numerous
Conclusion
Over the past several years, a number of exciting advances have been made in the field of nanobiodiagnostics largely due to advances in nanoparticle synthesis and the control over their surface binding properties. Specifically, the bio-barcode method has taken advantage of the properties of gold nanoparticles in order to generate a wide range of biomolecule detection assays that now rival or surpass the selectivity, sensitivity, and multiplexing capabilities of many conventional assays.
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The authors of this paper are associated with a company and hold patents, which pertain to some of the discoveries reported here.