Enzyme-free detection and quantification of double-stranded nucleic acids.
We have developed a fully enzyme-free SERRS hybridization assay for specific detection of double-stranded DNA sequences. Although all DNA detection methods ranging from PCR to high-throughput sequencing rely on enzymes, this method is unique for being totally non-enzymatic. The efficiency of enzymatic processes is affected by alterations, modifications, and/or quality of DNA. For instance, a limitation of most DNA polymerases is their inability to process DNA damaged by blocking lesions. As a result, enzymatic amplification and sequencing of degraded DNA often fail. In this study we succeeded in detecting and quantifying, within a mixture, relative amounts of closely related double-stranded DNA sequences from Rupicapra rupicapra (chamois) and Capra hircus (goat). The non-enzymatic SERRS assay presented here is the corner stone of a promising approach to overcome the failure of DNA polymerase when DNA is too degraded or when the concentration of polymerase inhibitors is too high. It is the first time double-stranded DNA has been detected with a truly non-enzymatic SERRS-based method. This non-enzymatic, inexpensive, rapid assay is therefore a breakthrough in nucleic acid detection.
Laboratoire de Géologie de Lyon, Terre, Planètes, Environnement, UMR 5276, Université Lyon 1, CNRS, Ecole Normale Supérieure de Lyon, 46 allée d'Italie, 69364, Lyon Cedex 07, France. cecile.feuillie@ens-lyon.org
Cécile
C
Feuillie
Maxime Mohamad
MM
Merheb
Benjamin
B
Gillet
Gilles
G
Montagnac
Catherine
C
Hänni
Isabelle
I
Daniel
D004247:DNA / Q000032:analysis*; D009696:Nucleic Acids / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods
D009696:Nucleic Acids; D004247:DNA