Nucleic Acids Research, 2002, Vol. 30, No. 2 e5
© 2002 Oxford University Press
A new class of homogeneous nucleic acid probes based on specific displacement hybridization
The Key Laboratory of Cell Biology and Tumor Cell Engineering of the Ministry of Education and 1Cancer Research Center, School of Life Sciences, Xiamen University, Xiamen 361005, Fujian, China
We have developed a new class of probes for homogeneous nucleic acid detection based on the proposed displacement hybridization. Our probes consist of two complementary oligodeoxyribonucleotides of different length labeled with a fluorophore and a quencher in close proximity in the duplex. The probes on their own are quenched, but they become fluorescent upon displacement hybridization with the target. These probes display complete discrimination between a perfectly matched target and single nucleotide mismatch targets. A comparison of double-stranded probes with corresponding linear probes confirms that the presence of the complementary strand significantly enhances their specificity. Using four such probes labeled with different color fluorophores, each designed to recognize a different target, we have demonstrated that multiple targets can be distinguished in the same solution, even if they differ from one another by as little as a single nucleotide. Double-stranded probes were used in real-time nucleic acid amplifications as either probes or as primers. In addition to its extreme specificity and flexibility, the new class of probes is simple to design and synthesize, has low cost and high sensitivity and is accessible to a wide range of labels. This class of probes should find applications in a variety of areas wherever high specificity of nucleic acid hybridization is relevant.
* To whom correspondence should be addressed at present address: Department of Molecular Genetics, Public Health Research Institute, Room 973, 455 First Avenue, New York, NY 10016, USA. Tel: +1 212 578 0858; Fax: +1 212 576 8471; Email: qing{at}phri.nyu.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Spandidos, X. Wang, H. Wang, and B. Seed PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification Nucleic Acids Res., November 11, 2009; (2009) gkp1005v1. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Huang, J. Salituro, N. Tang, K.-C. Luk, J. Hackett Jr, P. Swanson, G. Cloherty, W.-B. Mak, J. Robinson, and K. Abravaya Thermodynamically modulated partially double-stranded linear DNA probe design for homogeneous real-time PCR Nucleic Acids Res., August 13, 2007; 35(16): e101 - e101. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Huang, Y. Cheng, Q. Guo, and Q. Li Preparation of a chimeric armored RNA as a versatile calibrator for multiple virus assays. Clin. Chem., July 1, 2006; 52(7): 1446 - 1448. [Full Text] [PDF]
|
|
|
|
|
|
S. Narayanan, J. Gall, and C. Richert Clamping down on weak terminal base pairs: oligonucleotides with molecular caps as fidelity-enhancing elements at the 5'- and 3'-terminal residues Nucleic Acids Res., May 20, 2004; 32(9): 2901 - 2911. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Rajendran and A. D. Ellington In vitro selection of molecular beacons Nucleic Acids Res., October 1, 2003; 31(19): 5700 - 5713. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. E. Marras, F. R. Kramer, and S. Tyagi Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes Nucleic Acids Res., November 1, 2002; 30(21): e122 - e122. [Abstract] [Full Text] [PDF]
|
|