NON-ENZYMATIC LIPOSOME-LINKED CLOSELY SPACED ARRAY ELECTRODES ASSAY (NEL-ELA) FOR DETECTING AND QUANTIFYING NUCLEIC ACIDS

摘要

Target nucleic acids or amplicons thereof bound to immobilized capture oligonucleotides by molecular biological reations, are detected and quantified with affinity liposomes containing encapsulated electrochemically detectable reporter molecules susceptible to redox recycling and surface-attached affinity components capable of specifically binding to captured target nucleic acids or amplicons thereof in a structure restricted manner. Specifically bound affinity liposomes are lysed by temperature- or detergent-mediated mechanisms and released reporter molecules are quantitated via redox recycling using voltammetry in conjunction with a closely spaced array of thin film nobel metal electrodes. The quantitiy of released reporter molecules is a proportional measure of the quantity of target nucleic acids in the sample. For amplified assay procedures polymeric carrier molecules capable of binding multiple affinity liposomes or preformed complexes of affinity liposomes are utilized.