Development of a lipase-based optical assay for detection of DNA
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Author list: Pinijsuwan S., Shipovskov S., Surareungchai W., Ferapontova E.E., Gothelf K.V.
Publication year: 2011
Volume number: 9
Issue number: 18
Start page: 6352
End page: 6356
Number of pages: 5
ISSN: 1477-0520
eISSN: 1477-0520
Languages: English-Great Britain (EN-GB)
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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. ฉ 2011 The Royal Society of Chemistry.
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