Target substance detection method using aptamer
View Patent ↗The present invention relates to a method and kit of detecting a target material using an aptamer, and more particularly to a method and kit for detecting a target material, in which a sample and a second aptamer are added to a first aptamer immobilized on a solid phase so as to form a bond sandwiched between the first aptamer, the target material and the second aptamer, to an FET sensor-based method and kit for detecting a target material, and to an AAO sensor-based method and kit for detecting a target material. The inventive method for a target material using an aptamer can detect even low-molecular-weight materials which were difficult to detect in the prior art, thereby enabling detection of disease-related metabolites, environmental pollutants and food toxins in solutions. In addition, the detection method of the present invention is a direct and simple method and is highly cost-effective, because it uses the aptamer which can be consistently reproduced and can be produced at low costs. Thus, the present invention is very useful.
1. A method of detecting a small, apolar molecule using aptamers, the method comprising the steps of:
(a) adding a sample containing a small, apolar molecule, and a second aptamer binding specifically to the small, apolar molecule and having a label attached thereto, to a first aptamer immobilized on a solid phase and binding specifically to the small, apolar molecule, to form a mixture, and incubating the mixture; and
(b) analyzing the label to detect the small, apolar molecule.
2. The method of claim 1 , wherein the label includes fluorescent materials.
3. The method of claim 2 , wherein analysis of the fluorescent materials is performed by measuring the generated luminescence or color change of the fluorescent materials.
4. The method of claim 1 , wherein the sample is collected from one or more of water, blood, urine, tears, sweat, salvia, lymph, cerebrospinal fluids, soil, air, food, waste, animal and plant organs, and animal and plant tissues.
5. The method of claim 1 , wherein the first aptamer or the second aptamer is selected from aptamers represented by nucleic acid sequences of SEQ ID NOS: 2 to 28.
6. The method of claim 1 , wherein the small, apolar molecule is bisphenol A.
7. A method of detecting a small, apolar molecule using aptamers, the method comprising the steps of:
(a) adding a sample containing a small, apolar molecule, and a second aptamer binding specifically to the small, apolar molecule, to a first aptamer immobilized on a solid phase and binding specifically to the small, apolar molecule, to form a mixture, and incubating the mixture;
(b) binding a label to the second aptamer; and
(c) analyzing the label to detect the small, apolar molecule.
8. The method of claim 7 , wherein binding the label in step (b) is performed by complementarily binding a nucleic acid fragment to the second aptamer, in which the nucleic acid fragment has a label attached thereto and is capable of complementary binding to the second aptamer.
9. The method of claim 8 , wherein the nucleic acid fragment complementary bind to one end of the second aptamer.
10. The method of claim 7 , wherein the first aptamer or the second aptamer is selected from aptamers represented by nucleic acid sequences of SEQ ID NOS: 2 to 28.
11. The method of claim 7 , wherein the small, apolar molecule is bisphenol A.
12. A kit for detecting a small, apolar molecule, the kit comprising: a solid phase having immobilized thereon a first aptamer binding specifically to the small, apolar molecule; and a detection reagent containing a second aptamer binding specifically to the small, apolar molecule.
13. The kit of claim 12 , wherein the second aptamer has a label attached thereto.
14. The kit of claim 12 , wherein the kit further comprises a nucleic acid fragment having a label attached thereto and being capable of complementary binding to the second aptamer.
15. A kit for detecting bisphenol A, the kit comprising: a solid phase having immobilized thereon a first aptamer binding specifically to bisphenol A; and a detection reagent containing a second aptamer binding specifically to bisphenol A, wherein the first aptamer or the second aptamer is selected from aptamers represented by nucleic acid sequences of SEQ ID NOS: 2 to 28.
16. The method of claim 1 , wherein the label is a nucleic acid fragment and being capable of complementarily binding to the end of the aptamer.