title: Development of a lipase-based optical assay for detection of DNA.
abstract: A lipase-based assay for detection of specific DNA sequences has been developed. Lipase from Candida antarctica was conjugated to DNA and captured on magnetic beads in a sandwich assay, in which the binding was dependent on the presence of a specific target DNA. For amplification and to generate a detectable readout the captured lipase was applied to an optical assay that takes advantage of the enzymatic activity of lipase. The assay applies p-nitrophenol octanoate (NPO) as the substrate and in the presence of lipase the ester is hydrolyzed to p-nitrophenolate which has a strong absorbance at 405 nm. The method provides detection a detection limit of 200 fmol target DNA and it was able to distinguish single base mismatches from the fully complementary target.
authors.affiliation: Center for DNA Nanotechnology, Department of Chemistry and iNANO, Aarhus University, Aarhus C, Denmark.
authors.forename: Suttiporn
authors.initials: S
authors.lastname: Pinijsuwan
authors.forename: Stepan
authors.initials: S
authors.lastname: Shipovskov
authors.forename: Werasak
authors.initials: W
authors.lastname: Surareungchai
authors.forename: Elena E
authors.initials: EE
authors.lastname: Ferapontova
authors.forename: Kurt V
authors.initials: KV
authors.lastname: Gothelf
mesh_terms: D020137:Base Pair Mismatch; D015374:Biosensing Techniques / Q000379:methods*; D002175:Candida / Q000201:enzymology*; D004247:DNA / Q000032:analysis*; D005656:Fungal Proteins; D008049:Lipase / Q000378:metabolism*; D009596:Nitrophenols / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D005656:Fungal Proteins; D009596:Nitrophenols; D004247:DNA; D008049:Lipase; C431349:lipase B, Candida antarctica; C024836:4-nitrophenol