Dry reagent particle assay and device having multiple test zones and method therefor

Abstract

The present invention relates to a dry reagent assay device having at least one test zone and at least one reference zone, which provides an internal mechanism for assuring correct and reliable assay procedures and reagent qualities. In one embodiment, the present invention relates to an assay device having, at least one test zone for detecting at least one analyte in a sample by reacting the sample with a labeled indicator reagent, and a reference zone for receiving, unreacted labeled indicator reagent.

1 - 50 . (canceled) 
     
     
         51 . A device for determining the presence of an analyte in a sample, the device comprising: a bibulous member capable of being traversed by the sample; a first zone on the bibulous member for receiving and contacting the sample containing a diffusively immobilized, particle-linked antigen, the particle-linked antigen reacting in the presence of the analyte to form a mixture; a second zone on the bibulous member for receiving and contacting the mixture containing a non-diffusely immobilized antibody capable of binding the particle-linked antigen and free sample antigen, the second zone expresses a detectable response inversely related to the analyte level in the sample; a third zone on the bibulous member for receiving and contacting the remaining mixture containing a non-diffusively immobilized first member of a specific binding pair, capable of specifically binding to its specific binding partner which is the second member of the specific binding pair on the surface of the particle-linked antigen, which second member of the specific binding pair is not antigenically related to the sample antigen and will not effectively compete with the antigen to bind to an anti-antigen monoclonal antibody, the third zone expresses a detectable response related to the analyte level in the sample; and means for determining the analyte level in the sample from the detectable responses in the second and third zones. 
     
     
         52 . The device of  claim 51  wherein the determining means further comprises determining the sum of the detectable responses from the test zones to be within a pre-determined range of response. 
     
     
         53 . The device of  claim 51  wherein the determining means further comprises determining the level of analyte in one test zone by comparison to the total detectable response. 
     
     
         54 . The device of  claim 51  wherein the determining means further comprises determining the level of analyte using the detectable response in one test zone by comparison to a pre-determined standard. 
     
     
         55 . The device of  claim 51  wherein the determining means further comprises determining the level of analyte using the detectable response in one test zone in comparison to the detectable response in the other test zone. 
     
     
         56 . The device of  claim 51  wherein the determining means further comprises determining the level of analyte using the detectable response in one test zone in one portion of the pre-determined range of analyte and the detectable response in the other test zone in a different portion of the pre-determined range of analyte. 
     
     
         57 . A device for determining the presence of an analyte in a sample, the device comprising: a bibulous member capable of being traversed by the sample; a first zone on the bibulous member for receiving and contacting the sample containing a diffusively immobilized particle-linked antibody, the particle-linked antibody reacting in the presence of the analyte to form a conjugate included in a mixture; a second zone on the bibulous member for receiving and contacting the mixture containing a non-diffusively immobilized antigen capable of being bound by the particle-linked antibody, the second zone expresses a detectable response inversely related to the analyte level in the sample; a third zone on the bibulous member for receiving and contacting the remaining mixture containing a non-diffusively immobilized first member of a specific binding pair capable of specifically binding to its specific binding partner which is the second member of the specific binding pair on the surface of the particle-linked antigen, which second member of the specific binding pair is not antigenically related to the sample antigen so it will not effectively compete with the antigen to bind to an anti-antigen monoclonal antibody, the third zone expresses a detectable response related to the analyte level in the sample; and means for determining the analyte level in the sample from the detectable responses in the second and third zones. 
     
     
         58 . The device of  claim 57  wherein the determining means further comprises determining the sum of the detectable responses from the test zones is within a pre-determined range of response. 
     
     
         59 . The device of  claim 57  wherein the combining means further comprises determining the level of analyte in one zone by comparison to the total detectable response. 
     
     
         60 . The device of  claim 57  wherein the combining means further comprises determining the level of analyte using the detectable response in one test zone by comparison to a pre-determined standard. 
     
     
         61 . The device of  claim 57  wherein the combining means further comprises determining the level of analyte using the detectable response in one test zone in comparison to the detectable response in the other test zone. 
     
     
         62 . The device of  claim 57  wherein the combining means further comprises determining the level of analyte using the detectable response in one test zone in one portion of the pre-determined range of analyte and the detectable response in the other test zone in a different portion of the pre-determined range of analyte. 
     
     
         63 . A method for determining the level of at least one analyte in a sample, the method comprising the steps of; contacting the sample with an end portion of a bibulous strip having a plurality of zones; wicking the sample to a labeled indicator reagent diffusively immobilized on the bibulous strip; reacting the labeled indicator reagent in the presence of the analyte to form a mixture; wicking the mixture to a first reagent non-diffusely immobilized on the bibulous strip; reacting the first reagent in the presence of the mixture to form a first reaction product and a detectable response inversely related to the analyte level in the sample; wicking the remaining mixture to a second reagent non-diffusely immobilized on the bibulous strip; reacting the second reagent in the presence of the remaining mixture to form a second reaction product and a detectable response related to the analyte level in the sample; and determining the analyte level in the sample from the detectable responses in the reacting steps with the first and second reagents. 
     
     
         64 . The method of  claim 63  wherein: the reacting step with the labeled indicator reagent comprises forming a mixture including particle-linked antigen with the analyte; the reacting step with the first reagent comprises binding an antibody with the particle-linked antigen and the analyte; the reacting step with the second reagent comprises binding a first member of a specific binding pair to a second member of the specific binding pair on the particle-linked antigen, the second member of the specific binding pair is not a specific binding partner to the analyte. 
     
     
         65 . The method of  claim 63  wherein: the reacting step with the labeled indicator reagent comprises forming a mixture including a particle-linked antibody with the analyte; the reacting step with the first reagent comprises substantially binding the particle-linked antibody with an immobilized antigen; the reacting step for the second reagent comprises binding a first member of a specific binding pair to a second member of the specific binding pair on the particle-linked antibody, the second member of the specific binding pair is not a specific binding pair to the analyte.