Amperometric biosensor and detecting method using the same

A method for manufacturing an amperometric biosensor for sensing concentration of a target analyte, comprising:

• a) providing a plurality of insulator substrates each having an electrode unit disposed thereon, wherein the electrode unit includes a working electrode and a reference electrode spaced apart from the working electrode;
• b) disposing a blocking member on each of the insulator substrates in a manner that the reference electrode is not covered thereby and that the blocking member surrounds a portion of the working electrode, the blocking member being formed with an opening that exposes said portion of the working electrode, and cooperating with the insulator substrate to define a space;
• c) introducing a biorecognition element that is reactive to the target analyte into the space defined in step b), so as to dispose the biorecognition element on a surface of said portion of the working electrode that is exposed from the opening of the blocking member;
• and
• d) connecting electrically the electrode units to each other.

A method for detecting concentration of a target analyte, comprising:

• a) connecting electrically a plurality of sensing members to form an input of a detector, wherein each of the sensing members includes an insulator substrate and an electrode unit disposed on the insulator substrate, the electrode unit including a working electrode with a biorecognition element disposed thereon for reaction with the target analyte, and a reference electrode spaced apart from the working electrode;
• b) introducing the target analyte to each of the sensing members, so as to bring the target analyte into contact with at least a portion of the working electrode and at least a portion of the reference electrode of each of the sensing members;
• c) configuring the detector to provide a predetermined voltage between the working electrode and the reference electrode of each of the sensing members, so as to generate a current flowing through the target analyte introduced to each of the sensing members;
• and
• d) configuring the detector to detect the currents generated in step c) for subsequent concentration analysis of the target analyte.