Method for electrical detection of a binding reaction
First Claim
1. A method of detecting a substance in a test sample, which substance is a first one of a pair of first and second substances that undergo a specific binding reaction with each other, including the steps of:
- mixing the sample with particles under conditions effective to cause binding of the first substance, if present in the sample, to the surfaces of the particles;
contacting the particles having the first substance bound to the surfaces thereof with a layer of the second of the pair of substances, which layer forms a path between a pair of spaced-apart electrical conductors superposed on a substantially nonelectrically conductive base, the layer of the second substance being bound to the base, such that the binding reaction between the first and second substances causes the particles to be bound to the path in aggregates;
removing particles which are bound to the path as a result of non-specific binding and particles which are unbound to the path;
coating exposed outer surfaces of the aggregates with an electrically conductive substance that adheres selectively to the particles forming the aggregates but which does not adhere to the remainder of the path;
removing electrically conductive substances which remains unadhered to the aggregates;
connecting each of the conductors to an electrical circuit which includes a source of electrical energy, the conductors and the path therein; and
measuring a change in the electrical current flow through the circuit caused by formation of the coated aggregates on the path, the electrical change indicating the level of the substance to be detected in the test sample.
12 Assignments
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Accused Products
Abstract
A method is disclosed for detecting the occurrence of a binding or complex-forming reaction between specific substances by utilizing the binding reaction to modify an electrical circuit, and then measuring a change in the electrical state of the circuit. A diagnostic element useful in such a method includes a layer of a biogenic substance, such as an antigen, coated onto a non-conductive base between a pair of electrical conductors superposed on the base. Antibodies which react with the antigen are treated so that they become bound to particles. The particles having antibody bound thereto are then added to is the antigen lay e base and allowed to react therewith. The particles are thereby bound to the base due to the binding reaction between the antigen and antibody to thereby form aggregates of electrically conductive particles which modify the circuit. The particles are then selectively coated with a conductive substance. The method of the invention is highly useful for the detection of antigens and antibodies in the blood serum of a human patient.
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Citations
20 Claims
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1. A method of detecting a substance in a test sample, which substance is a first one of a pair of first and second substances that undergo a specific binding reaction with each other, including the steps of:
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mixing the sample with particles under conditions effective to cause binding of the first substance, if present in the sample, to the surfaces of the particles; contacting the particles having the first substance bound to the surfaces thereof with a layer of the second of the pair of substances, which layer forms a path between a pair of spaced-apart electrical conductors superposed on a substantially nonelectrically conductive base, the layer of the second substance being bound to the base, such that the binding reaction between the first and second substances causes the particles to be bound to the path in aggregates; removing particles which are bound to the path as a result of non-specific binding and particles which are unbound to the path; coating exposed outer surfaces of the aggregates with an electrically conductive substance that adheres selectively to the particles forming the aggregates but which does not adhere to the remainder of the path; removing electrically conductive substances which remains unadhered to the aggregates; connecting each of the conductors to an electrical circuit which includes a source of electrical energy, the conductors and the path therein; and measuring a change in the electrical current flow through the circuit caused by formation of the coated aggregates on the path, the electrical change indicating the level of the substance to be detected in the test sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of detecting a substance in a test sample, which substance is a first one of a pair of first and second substances that undergo a specific binding reaction with each other, including the steps of:
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mixing the sample with particles having the second one of the substances bound to the surfaces thereof under conditions effective to cause binding of the first substance, if present in the sample, to the second substance on the surfaces of the particles; contacting the particles with a layer of the first substance, which layer forms a path between a pair of spaced-apart electrical conductors superposed on a substantially nonelectrically conductive base, the layer of the first substance being bound to the base, such that the binding reaction between the first and second substances causes the particles to be bound to the path in aggregates, and particles on which the second substance became bound to the first substance from the test sample in sufficient quantity do not become bound to the path; removing particles which are bound to the path as a result of non-specific binding and particle which are unbound to the path; coating exposed outer surfaces of the aggregates with an electrically conductive substance that adheres selectively to the particles forming the aggregates but which does not adhere to the remainder of the path; removing electrically conductive substance which remains unadhered to the aggregates; connecting each of the conductors to an electrical circuit which includes a source of electrical energy, the conductors and the path therein; and measuring a change in the electrical current flow through the circuit caused by formation of the coated aggregates on the path, the electrical change indicating the level of the substance to be detected in the test sample. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification