Terbium fluorescence as a sensitive, inexpensive probe for UV-induced damage in nucleic acids

• Author(s) / Creator(s)

  • El-Yazbi, Amira F.
  • Loppnow, Glen R.

• Much effort has been focused on developing methods for detecting damaged nucleic acids. However, almost all of the proposed methods consist of multi-step procedures, are limited, require expensive instruments, or suffer from a high level of interferences. In this paper, we present a novel simple, inexpensive, mix-and-read assay that is generally applicable to nucleic acid damage and uses the enhanced luminescence due to energy transfer from nucleic acids to terbium(III) (Tb3+). Single-stranded oligonucleotides greatly enhance the Tb3+ emission, but duplex DNA does not. With the use of a DNA hairpin probe complementary to the oligonucleotide of interest, the Tb3+/hairpin probe is applied to detect ultraviolet (UV)-induced DNA damage. The hairpin probe hybridizes only with the undamaged DNA. However, the damaged DNA remains single-stranded and enhances the intrinsic fluorescence of Tb3+, producing a detectable signal directly proportional to the amount of DNA damage. This allows the Tb3+/hairpin probe to be used for sensitive quantification of UV-induced DNA damage. The Tb3+/hairpin probe showed superior selectivity to DNA damage compared to conventional molecular beacons probes (MBs) and its sensitivity is more than 2.5 times higher than MBs with a limit of detection of 4.36 ± 1.2 nM. In addition, this probe is easier to synthesize and more than eight times cheaper than MBs, which makes its use recommended for high-throughput, quantitative analysis of DNA damage.

• Date created

  2013-01-01

• Subjects / Keywords

  • Nucleic acid damage
  • Fluorescence sensor
  • Terbium
  • Fluorescence
  • Hairpin probe

• Type of Item

  Article (Draft / Submitted)

• DOI

  https://doi.org/10.7939/r3-jk7m-et42

• License

  © 2013 Elsevier B.V. All rights reserved.