Method of detecting target material, sensor chip, and detecting device
A method of detecting a target material ( 3 ) includes: a step of preparing a complex ( 11 ), the complex including an aptamer ( 1 ) to which a target material ( 3 ) in a specimen specifically binds, a first nucleic acid fragment ( 2 ) that has a base sequence complementary to the aptamer ( 1 ), and a fixing member ( 4 ) to which a part of the aptamer ( 1 ) and a part of the first nucleic acid fragment ( 2 ) are fixed, in which the aptamer ( 1 ) has a double strand-forming site ( 5 ) capable of forming a double strand with the first nucleic acid fragment ( 2 ); a step of separating the first nucleic acid fragment 92 ) from the double strand-forming site ( 5 ) of the aptamer ( 1 ) by binding the target material ( 3 ) to the aptamer ( 1 ); and a step of detecting the cleavage of the double strand.
1. A method of detecting a target material, the method comprising:
a step of preparing a complex, the complex including
an aptamer to which a target material in a specimen specifically binds,
a first nucleic acid fragment that has a base sequence complementary to the aptamer, and
a fixing member to which a part of the aptamer and a part of the first nucleic acid fragment are fixed,
in which the aptamer has a double strand-forming site capable of forming a double strand with the first nucleic acid fragment;
a step of separating the first nucleic acid fragment from the double strand-forming site of the aptamer by binding the target material to the aptamer; and
a step of detecting cleavage of the double strand where the first nucleic acid fragment is separated from the aptamer;
wherein a base sequence of the aptamer to which the target material binds comprises at least a part of a base sequence of a double strand-forming site formed between the aptamer and the first nucleic acid fragment.
2. The method of detecting a target material according to claim 1 ,
wherein the double strand-forming site of the aptamer has only a base sequence complementary to the base sequence of the first nucleic acid fragment.
3. The method of detecting a target material according to claim 1 ,
wherein the complex includes a linking portion through which a part of the aptamer and a part of the first nucleic acid fragment are linked to each other, and
the linking portion is fixed to the fixing member.
4. The method of detecting a target material according to claim 1 ,
wherein the step of detecting the cleavage of the double strand includes
a step of binding a second nucleic acid fragment, which has a base sequence complementary to the first nucleic acid fragment, to the first nucleic acid fragment which is separated from the double strand-forming site, and
a step of detecting a bond between the first nucleic acid fragment and the second nucleic acid fragment to detect the cleavage of the double strand.
5. The method of detecting a target material according to claim 4 ,
wherein the fixing member has a solvophilic region in a different region from a fixing region where the aptamer and the first nucleic acid fragment are fixed.
6. The method of detecting a target material according to claim 4 ,
wherein the step of binding a second nucleic acid fragment includes a step of binding the second nucleic acid fragment, which is fixed to a substrate, to the first nucleic acid fragment which is separated from the double strand-forming site, and
the substrate has a solvophilic region in a different region from a fixing region where the second nucleic acid fragment is fixed.
7. The method of detecting a target material according to claim 1 , further comprising:
a cleavage energy applying step of applying a cleavage energy, which is higher than or equal to a binding energy of a double strand between the aptamer and the first nucleic acid fragment, before or during the step of separating the first nucleic acid fragment from the double strand-forming site of the aptamer; and
a double strand stabilizing step of strengthening a binding strength of a double strand between the aptamer and the first nucleic acid fragment, before or during the step of detecting the cleavage of the double strand;
wherein the cleavage energy applying step comprises heating a measurement solution.
8. The method of detecting a target material according to claim 1 , further comprising:
a cleavage energy applying step of applying a cleavage energy, which is lower than a binding energy of a double strand between the aptamer and the first nucleic acid fragment, before or during the step of separating the first nucleic acid fragment from the double strand-forming site of the aptamer; and
a double strand stabilizing step of strengthening a binding strength of a double strand between the aptamer and the first nucleic acid fragment, before or during the step of detecting the cleavage of the double strand;
wherein the double strand stabilizing step comprises cooling a measurement solution.