Luminescent Probes for Ultrasensitive Detection of Nucleic Acids

Novel amino-reactive derivatives of lanthanide-based luminescent labels of enhanced brightness and metal retention were synthesized and used for the detection of cDNA oligonucleotides by molecular beacons. Time-resolved acquisition of the luminescent signal that occurs upon hybridization of the prob...

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Bibliographic Details
Published in:	Bioconjugate chemistry 2010-02, Vol.21 (2), p.319-327
Main Authors:	Krasnoperov, Lev N, Marras, Salvatore A. E, Kozlov, Maxim, Wirpsza, Laura, Mustaev, Arkady
Format:	Article
Language:	eng
Subjects:	Absorption Amino acids Base Sequence Biopolymers Chelating Agents - chemistry Chemical compounds Chemical synthesis Deoxyribonucleic acid DNA Edetic Acid - analogs & derivatives Edetic Acid - chemistry Kinetics Lanthanoid Series Elements - chemistry Luminescence Luminescent Agents - chemical synthesis Luminescent Agents - chemistry Molecular Probes - chemistry Molecular Probes - metabolism Molecules Nucleic Acids - analysis Nucleic Acids - genetics Nucleic Acids - metabolism Pentetic Acid - chemistry Reproducibility of Results Spectrometry, Fluorescence
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Summary:	Novel amino-reactive derivatives of lanthanide-based luminescent labels of enhanced brightness and metal retention were synthesized and used for the detection of cDNA oligonucleotides by molecular beacons. Time-resolved acquisition of the luminescent signal that occurs upon hybridization of the probe to the target enabled the avoidance of short-lived background fluorescence, markedly enhancing the sensitivity of detection, which was less than 1 pM. This value is about 50 to 100 times more sensitive than the level achieved with conventional fluorescence-based molecular beacons, and is 10 to 60 times more sensitive than previously reported for other lanthanide-based hybridization probes. These novel luminescent labels should significantly enhance the sensitivity of all type of nucleic acid hybridization probes, and could dramatically improve the detection limit of other biopolymers and small compounds that are used in a variety of biological applications.
ISSN:	1043-1802 1520-4812