Disposable test strips with integrated reagent/blood separation layer

A method for forming a disposable test strip for use in a test meter which receives a disposable test strip and a sample of blood and performs an electrochemical analysis of the amount of a blood analyte in the sample, comprising:

• (a) forming first and second conductive elements on a substrate;
• (b) forming a layer of insulation covering the first conductive element, said layer of insulation having a first aperture therein aligned with a portion of the first conductive element in a sample application region;
• and
• (c) forming a integrated reagent/blood separation layer layer disposed on the insulation layer and making contact with the first conductive element through the first aperture in the insulation layer, said integrated reagent/blood separation layer comprising reagents for the electrochemical detection of glucose dispersed in a non-conductive matrix effective to exclude blood cells from the surface of the first conductive element while permitting access to the first conductive species by soluble electroactive species, whereby the first conductive element is isolated from direct contact with a sample placed on the test strip.