Method and system for nucleic acid detection using electroconductive or electrochemically active labels
AssigneeThe Hong Kong University of Science and Technology
InventorsI-Ming HSING, Ming Hung Thomas LEE, Xiaoteng LUO
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Abstract
A method for electrochemically or electrically detecting nucleic acids, utilizes electrochemically active or electrically conductive reporter materials. An electric voltage is applied and electric signals are measured to the electrodes that are suitable for detecting or quantifying the nucleic acid(s) in a sample. This technique is suitable for point-of-use applications, e.g. detecting bioanalytes in remote locations. A microchip, device, kit used adapted to be used for this method is also disclosed.
Claims15
A microchip comprising an electrochemically or electrically conductive electrode(s) provided on a surface adapted to receive a molecule comprising a target nucleic acid(s);
- wherein said microchip is adapted to be used for a solution phase method for electrochemically or electrically detecting target nucleic acid(s), said method comprising:
- a. providing a sample comprising:
- i. a first probe(s) having the same electrical as that of an electrode surface such that said first probe(s) is(are) repelled from said electrode surface;
- and
- ii. a second probe(s) comprising an electrochemically or electrically conductive labeled marker(s) coupled to an electrically neutral molecule(s), said second probe(s) being operatively linked to said first probe(s);
- b. providing said target nucleic acid(s) to said sample wherein said first probe(s) is(are) complementary to at least a portion of said target nucleic acid(s);
- c. applying an electric potential to said sample and detecting or measuring a signal(s) produced by said labeled marker(s);
- and
- wherein when first said probe(s) is(are) hybridized into said target nucleic acid(s), said second probe(s) is(are) released from said first probe(s) and said labeled marker(s) is(are) attracted to said electrode surface to produce a signal intensity change(s) when subjected to an electric potential, and such signal intensity change(s) correspond(s) to said target nucleic acid(s) in said sample.
A microchip comprising an electrochemically or electrically conductive electrode(s) provided on a surface adapted to receive a molecule comprising a nucleic acid(s);
- wherein said microchip is adapted to be used for a solid phase method for electrochemically or electrically monitoring or quantifying the amount of nucleic acid(s), said method comprising:
- a. contacting a sample comprising a target nucleic acid(s), a solid surface bound probe(s) comprising a first primer(s) provided with a sequence(s) that is complementary to at least a portion of one end of the target nucleic acid(s), a second primer(s) in solution that is complementary to at least a portion of the opposing end of the complementary strand of the target nucleic acid(s), and an electrochemically or electrically conductive labeled marker(s) that is(are) adapted for incorporation into a polynucleic acid(s) by chain polymerization and when incorporated thereof produces a signal(s) change(s) if subjected to an electric potential;
- b. adding a polymerase chain reaction enzyme(s) under conditions effective for PCR amplification to occur;
- c. applying an electric potential to the sample and detecting or measuring in real time an electric signal(s) produced by the labeled marker(s) incorporated into said solid surface bound probe(s);
- and
- d. quantifying the amount of nucleic acid(s) present in the sample and the amount of polynucleic acid(s) produced by correlating the change(s) in signal(s) over time with the formation of polynucleic acid(s).
- wherein said electric signal(s) is(are) detected or measured with a conductive electrode(s) placed in said sample;
- said electrode(s) is(are) treated with surface passivation.
Dependencies are inferred from the claim text.