Rapid, single-step nucleic acid detection

Kyle A. Cissell, Sean Campbell, Sapna K. Deo

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

A rapid detection method for nucleic acid based on bioluminescence resonance energy transfer (BRET) from the luminescence donor Renilla luciferase to an acceptor quantum dot upon oligonucleotide probe hybridization has been developed. Utilizing a competitive assay, we detected the target nucleic acid by correlating the BRET signal with the amount of target present in the sample. This method allows for the detection of as little as 4 pmol (20 nM) of nucleic acid in a single-step, homogeneous format both in vitro in a buffer matrix as well as in a cellular matrix. Using this method, one may perform nucleic acid detection in as little as 30 min, showing much improvement over time-consuming blotting methods and solid-phase methods which require multiple wash steps to remove unbound probe. This is the first report on the use of quantum dots as a BRET acceptor in the development of a nucleic acid hybridization assay.

Original languageEnglish (US)
Pages (from-to)2577-2581
Number of pages5
JournalAnalytical and bioanalytical chemistry
Volume391
Issue number7
DOIs
StatePublished - Aug 1 2008
Externally publishedYes

Keywords

  • Bioluminescence resonance energy transfer
  • Nucleic acid detection
  • Nucleic acid hybridization
  • Quantum dots
  • Renilla luciferase

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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