Electrochemical method to measure dna attachment to an...

Details Electrochemical method to measure dna attachment to an... Electrochemical method to measure dna attachment to an...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2524263

The present disclosure provides methods and compositions for conducting an assay to detect nucleic acid hybridization in the presence of oxygen. In particular, ruthenium complexes having a reduction potential that does not coincide with the reduction potential of molecular oxygen are disclosed and amperometric techniques for their use are described. In preferred embodiments, the ruthenium complex is ruthenium (III) pentaamine pyridine and the nucleic acid hybridization event that is detected is DNA hybridization. Further, techniques for enhancing detectable contrast between hybridized and unhybridized nucleic acids are disclosed. In particular, the use of elongated target strands as well as the use of uncharged probe strands are discussed.

La présente invention concerne des procédés et des compositions permettant de réaliser un essai visant à détecter une hybridation d'acides nucléiques en présence d'oxygène. Plus particulièrement, l'invention concerne des complexes de ruthénium présentant un potentiel de réduction ne coïncidant pas avec le potentiel de réduction de l'oxygène moléculaire, ainsi que des techniques ampérométriques destinées à leur utilisation. Dans des modes de réalisation préférés, le complexe de ruthénium se présente sous forme de ruthénium (III) penta-amine pyridine et l'événement d'hybridation d'acides nucléiques détecté est l'hybridation d'ADN. L'invention concerne en outre des techniques destinées à augmenter le contraste détectable entre des acides nucléiques hybridés et non hybridés. Plus particulièrement, elle se rapporte à l'utilisation de brins cibles allongés ainsi qu'à l'utilisation de brins sondes non chargés.

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