IP Library Patent Application 16302346
Patent Application
App. No. 16/302,346

METHOD FOR DETECTION OF A PCR PRODUCT

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Quick Facts
Patent No.
US None
App. No.
16/302,346
Abstract

A method for detecting a nucleic acid molecule in a biological sample includes amplifying a nucleic acid molecule to generate an amplicon having a single 5′-tail and coupling the 5′-tail to one of a plurality of capture probes on a surface of a sensor. The amplicon is converted to a single strand molecule and a target-specific catalyst cluster is bound to the single strand molecule. The catalyst cluster is subjected to metallization in order to detect the target nucleic acid.

Claims (26)

1 . A method for detecting a target nucleic acid molecule in a sample with a sensor comprising a first electrode and a second electrode coupled to a sensor surface in a spaced apart arrangement and a plurality of capture probes coupled to the sensor surface between the first electrode and the second electrode, the method comprising:

performing nucleic acid molecule amplification via polymerase chain reaction (PCR) using a first primer having a 5′-tail and a second primer having no 5′-tail to form a plurality of double-stranded amplicons having a first strand with a 5′-tail and a second strand with no tail;

hybridizing the plurality of amplicons to the plurality of capture probes;

converting the plurality of amplicons to a plurality of single strand molecules;

binding a catalyst cluster to an interior section of each of the plurality of single strand molecules;

contacting the plurality of single strand molecules having a catalyst cluster bound thereon with a metal or metal alloy to deposit the metal or metal alloy on the catalyst cluster; and

determining if an electrical current can be carried between the electrodes, the electrical current between the electrodes indicating presence of the target nucleic acid molecule in the sample.

2 . The method of claim 1 , wherein converting the plurality of amplicons to the plurality of single strand molecules comprises employing an exonuclease to digest the second strand with no tail.

3 . The method of claim 1 , wherein the catalyst cluster comprises a catalyst gold cluster and wherein the metal comprises gold.

4 . The method of claim 1 , further comprising forming a target specific catalyst cluster configured to bind to an interior region of the first strand of the amplicon.

5 . The method of claim 1 , wherein the catalyst cluster is a generic cluster having an adaptor oligonucleotide coupled thereto.

6 . The method of claim 5 , wherein the generic cluster has a plurality of oligonucleotides coupled thereto, each oligonucleotide configured to target a different amplicon.

7 . The method of claim 1 , wherein binding the catalyst cluster comprises binding a plurality of catalyst clusters to interior sections of each of the single strand molecules.

8 . A method for preparing a nucleic acid molecule detector comprising a first electrode and a second electrode coupled to a sensor surface in a spaced apart arrangement and a plurality of capture probes coupled to the sensor surface between the first electrode and the second electrode, the method comprising:

receiving a biological sample;

amplifying a nucleic acid molecule within the biological sample to generate an amplicon having a single 5′-tail;

coupling the 5′-tail of the amplicon to one of the plurality of capture probes;

employing an exonuclease to digest one strand of the amplicon to convert the amplicon to a single strand molecule;

synthesizing a target-specific catalyst cluster; and

contacting the catalyst cluster with the single strand molecule of the amplicon to bind the target-specific catalyst cluster to an interior region of the single strand molecule.

9 . The method of claim 8 , wherein the catalyst cluster comprises a catalyst gold cluster.

10 . The method of claim 8 , wherein synthesizing the target-specific catalyst cluster comprises hybridizing a base generic cluster with an adaptor oligonucleotide to form a catalyst cluster having a plurality of adaptor oligonucleotides coupled thereto.

11 . The method of claim 10 , wherein the adaptor oligonucleotide has a cluster binding sequence and an amplicon-specific binding sequence.

12 . The method of claim 10 , wherein hybridizing the base generic cluster comprises hybridizing the generic cluster with a plurality of adaptor oligonucleotides, each adaptor oligonucleotide configured to target a different amplicon.

13 . The method of claim 8 , wherein amplifying the nucleic acid molecule comprises performing polymerase chain reaction (PCR) using a first primer having a 5′-tail and a second primer having no 5′-tail.

14 . The method of claim 8 , wherein each of the plurality of capture probes comprises a functionalized oxide surface configured to immobilize molecules to the sensor surface.

Assignments (4)
CORRECTIVE ASSIGNMENT TO CORRECT THE THE 9TH PATENT NUMBER AND THE NATURE OF CONVEYANCE PREVIOUSLY RECORDED AT REEL: 60224 FRAME: 860. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT. Recorded Dec 13, 2024
From: INTEGRATED NANO-TECHNOLOGIES, INC.
To: CONNOLLY, DENNIS MICHAEL
Reel/Frame 069685/0794 →
COURT ORDER Recorded May 20, 2023
From: INTEGRATED NANO-TECHNOLOGIES, INC.
To: ENPLAS AMERICA, INC.
Reel/Frame 063710/0198 →
SECURITY INTEREST Recorded Jun 16, 2022
From: INTEGRATED NANO-TECHNOLOGIES, INC.
To: CONNOLLY, DENNIS MICHAEL
Reel/Frame 060224/0860 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 28, 2019
From: MURANTE, RICHARD S.; TANNOUS, VERA
To: INTEGRATED NANO-TECHNOLOGIES, INC.
Reel/Frame 048468/0951 →