Enzyme amplified electrochemical DNA detection
First Claim
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1. An oligonucleotide probe for enzyme amplified electrochemical target DNA detection, the oligonucleotide probe comprising:
- a site-specific sequence; and
a co-factor conjugated to the site-specific sequence or a mediator conjugated to the site-specific sequence;
wherein the co-factor is adapted to produce an enzymatic signal that is electrochemically detectable and the mediator is adapted to enhance an enzymatic signal that is electrochemically detectable.
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Abstract
An oligonucleotide probe for DNA detection is disclosed. The oligonucleotide probe includes a site-specific sequence. Either a co-factor or a mediator is conjugated to the site-specific sequence. The co-factor is adapted to produce an enzymatic signal that is electrochemically detectable. The mediator is adapted to enhance an enzymatic signal that is electrochemically detectable.
10 Citations
37 Claims
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1. An oligonucleotide probe for enzyme amplified electrochemical target DNA detection, the oligonucleotide probe comprising:
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a site-specific sequence; and
a co-factor conjugated to the site-specific sequence or a mediator conjugated to the site-specific sequence;
wherein the co-factor is adapted to produce an enzymatic signal that is electrochemically detectable and the mediator is adapted to enhance an enzymatic signal that is electrochemically detectable. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method of detecting target DNA in a sample, the method comprising:
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performing a DNA amplification process on the sample;
exposing an oligonucleotide probe to exonuclease activity, the oligonucleotide probe including a site-specific sequence and a co-factor conjugated to the site-specific sequence or a mediator conjugated to the site-specific sequence, the exposing thereby releasing a probe fragment including the co-factor or the mediator;
combining the probe fragment with an apo-enzyme or a holo-enzyme, wherein combining the probe fragment having co-factor with the apo-enzyme produces an enzymatic signal that is electrochemically detectable or wherein combining the probe fragment having the mediator with the holo-enzyme enhances an enzymatic signal that is electrochemically detectable; and
electrochemically detecting the enzymatic signal, thereby detecting the target DNA. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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30. A diagnostic device for detecting target DNA, comprising:
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at least one electrode having an apo-enzyme or a holo-enzyme immobilized thereon; and
an oligonucleotide probe in electrochemical contact with the at least one electrode, the probe including;
a site-specific sequence; and
one of a co-factor conjugated to the site-specific sequence and a mediator conjugated to the site-specific sequence;
wherein the co-factor or the mediator is adapted to release from the probe to form a probe fragment;
wherein the probe fragment having the co-factor binds to the apo-enzyme to produce an enzymatic signal that is electrochemically detectable;
or wherein the probe fragment having the mediator reacts with the holo-enzyme to enhance an enzymatic signal that is electrochemically detectable. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37)
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Specification