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Highly sensitive and selective oligonucleotide sensor for sickle cell disease gene using photon upconverting nanoparticles.

ABSTRACT: We report the design of an oligonucleotide sensor for the detection of point mutation associated with sickle cell disease. The sensor was based on luminescence resonance energy transfer between a donor and an acceptor. Photon upconverting nanoparticles (NaYF(4) doped with Yb(3+) and Er(3+)) were used as the donor and a conventional fluorophore, N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA), as the acceptor. The sensor could detect the perfectly matched target, in the background of the mismatched target or other oligonucleotides of random sequences. The detection limit of this sensor towards perfectly matched target was calculated to be 120 femtomoles, with no photobleaching. Oligonucleotide sensors of such design demonstrate high sensitivity and specificity.

SUBMITTER: Kumar M

PROVIDER: S-EPMC2753611 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature

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Highly sensitive and selective oligonucleotide sensor for sickle cell disease gene using photon upconverting nanoparticles.

Kumar Manoj M Guo Yanyan Y Zhang Peng P

Biosensors & bioelectronics 20080826 5

We report the design of an oligonucleotide sensor for the detection of point mutation associated with sickle cell disease. The sensor was based on luminescence resonance energy transfer between a donor and an acceptor. Photon upconverting nanoparticles (NaYF(4) doped with Yb(3+) and Er(3+)) were used as the donor and a conventional fluorophore, N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA), as the acceptor. The sensor could detect the perfectly matched target, in the background of the mismatc ...[more]

PMID: 18823772

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Project description:A typical lock-and-key sensing strategy, relying only on the most dominant interactions between the probe and target, could be too limiting. In reality, the information received upon sensing is much richer. Non-specific events due to various intermolecular forces contribute to the overall received information with different degrees, and when analyzed, could provide a much more powerful detection opportunity. Here, we have assembled a highly selective universal sensor array using water-soluble two-dimensional nanoparticles (nGO, MoS2 and WS2) and fluorescent DNA molecules. The array is composed of 12 fluorescently silent non-specific nanoreceptors (2D-nps) and used for the identification of three radically different systems; five proteins, three types of live breast cancer cells and a structure-switching event of a macromolecule. The data matrices for each system were processed using Partial Least Squares (PLS) discriminant analysis. In all of the systems, the sensor array was able to identify each object or event as separate clusters with 95% confidence and without any overlap. Out of 15 unknown entities with unknown protein concentrations tested, 14 of them were predicted successfully with correct concentration. 8 breast cancer cell samples out of 9 unknown entities from three cell types were predicted correctly. During the assembly of each nanoprobe, the intrinsic non-covalent interactions between unmodified 2D nanoparticles and ssDNAs were exploited. The unmodified 2D materials offer remarkable simplicity in the layout and the use of ssDNAs as probes provides limitless possibilities because the natural interaction of a ssDNA and 2D surface can be fine-tuned with the nucleobase composition, oligonucleotide length and type of 2D nanomaterial. Therefore, the approach described here can be advanced and fine-tuned indefinitely for meeting a particular sensing criterion. Though we have only studied three distinct elements, this approach is universal enough to be applied to a wide-range of systems.

Dataset Information

Highly sensitive and selective oligonucleotide sensor for sickle cell disease gene using photon upconverting nanoparticles.

Publications

Highly sensitive and selective oligonucleotide sensor for sickle cell disease gene using photon upconverting nanoparticles.

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