Highly sensitive colorimetric enzyme-linked oligonucleotide assay based on cyclodextrin-modified polymeric surfaces.
In this paper, we describe the development of an enzyme-linked oligonucleotide assay for the detection of a human leukocyte antigen allele associated with celiac disease based on cyclodextrin-modified polymeric surfaces. The surface of maleimide-pre-coated plates was modified with a layer of thiolated cyclodextrin polymer and used for the supramolecular capture of adamantane or ferrocene-modified carboxymethylcellulose polymers bearing DNA probes. The assay was optimised in terms of incubation time, temperature, and surface chemistry and applied to the highly sensitive and selective detection of HLA sequences with a limit of detection of 0.7 nM. A real sample analysed using this platform showed an excellent correlation with maleimide-activated plates using thiolated DNA probes.
Departament d'Enginyeria Química, Universitat Rovira i Virgili, Tarragona, Spain.
Mayreli
M
Ortiz
Mabel
M
Torréns
Alex
A
Fragoso
Ciara K
CK
O'Sullivan
D001483:Base Sequence; D003124:Colorimetry / Q000379:methods*; D003505:Cyclodextrins / Q000737:chemistry*; D015342:DNA Probes; D057230:Limit of Detection; D009841:Oligonucleotides / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D003505:Cyclodextrins; D015342:DNA Probes; D009841:Oligonucleotides; D011108:Polymers