Method and devices for rapid diagnosis of foot-and-mouth disease

Abstract

A rapid immunoassay method and apparatus for detecting foot and mouth disease virus are disclosed. The method and test device permit pen-side testing of animals and provide test results within a relatively short time period. In a preferred embodiment, the method and apparatus provide a means for differentiating between FMDV-infected and FMDV-vaccinated animals.

1 - 7 . (canceled) 
     
     
         8 . A method for simultaneously detecting and differentiating between analytes comprising antibodies to FMDV structural proteins and antibodies to FMDV non-structural proteins in a sample, comprising:
 a. obtaining a fluid test sample to be assayed for said antibodies of interest;   b. supplying a rapid measurement device comprising a membrane strip with a proximal end containing a labeled binding partner and a labeled control reagent, and a distal end containing a zone having at least one immobilized capture reagent specific for an FMDV structural protein comprising at least one of VP1, VP2, VP3 or VP4, and a second zone having at least one immobilized capture reagent specific for an FMDV non-structural protein comprising at least one of Lb, 2B, 2C, 3A, 3AB, or 3ABC, and a third band of immobilized control capture reagent wherein said fluid test sample moves from said proximal end to said distal end by capillary action;   c. contacting said membrane strip with said fluid test sample;   d. maintaining said membrane strip under conditions which are sufficient to allow said analyte of interest to bind to said labeled binding partner, forming a specific binding complex, and to allow fluid to transport said specific binding complex and said labeled control reagent in said fluid by capillary action through said membrane strip to said immobilized capture reagent specific for an FMDV structural protein and said adjacent immobilized capture reagent specific for an FMDV non-structural protein and to said immobilized control capture reagent;   e. maintaining said membrane strip under conditions which are sufficient to allow said specific binding complex in said fluid to bind to at least one of said immobilized capture reagent specific for an FMDV structural protein and/or to bind to at least one of said immobilized capture reagent specific for an FMDV non-structural protein and said labeled control reagent to bind to said immobilized control capture reagent;   f. detecting an amount of said analyte of interest in said fluid test sample and an amount of internal control signal, as indicated by an intensity of a signal producing system; and   g. differentiating between said antibodies to FMDV structural proteins and said antibodies to FMDV non-structural proteins.   
     
     
         9 . The method of  claim 8  wherein said labeled binding partner is at least one of VP1, 3D, 2C or 3ABC and at least one of VP1, VP2, VP3 and VP4. 
     
     
         10 . The method of  claim 8  wherein said labeled binding partner is protein G and/or protein A. 
     
     
         11 . The method of  claim 8  wherein said membrane strip comprises an application point comprising a filter pad containing at least one of said labeled binding partner and said labeled control reagent. 
     
     
         12 . A method for quantitatively measuring an amount of an analyte of interest in a sample for differentiating FMDV infection from FMDV vaccination, comprising:
 a. obtaining a fluid test sample to be assayed for said analyte of interest;   b. supplying a rapid measurement device comprising a membrane strip with a proximal end containing a labeled binding partner and a labeled control reagent, and a distal end containing a band of an immobilized capture reagent comprising at least one of VP1, 3D, 2C or 3ABC, and a second band of immobilized control capture reagent, wherein said fluid test sample moves from said proximal end to said distal end by capillary action;   c. contacting said membrane strip with said fluid test sample;   d. maintaining said membrane strip under conditions which are sufficient to allow said analyte of interest to bind to said labeled binding partner, forming a specific binding complex, and to allow fluid to transport said specific binding complex and said labeled control reagent in said fluid by capillary action through said membrane strip to said immobilized capture reagent and said immobilized control capture reagent;   e. maintaining said membrane strip under conditions which are sufficient to allow said specific binding complex in said fluid to bind to said immobilized capture reagent and said labeled control reagent to bind to said immobilized control capture reagent; and   f. quantitating an amount of said analyte of interest in said fluid test sample as indicated by an intensity of a signal producing system generated in said fluid test sample relative to an intensity of signals generated in a standard curve.   
     
     
         13 . The method of  claim 12  wherein said labeled binding partner is at least one of VP1, 3D, 2C or 3ABC. 
     
     
         14 . The method of  claim 12  wherein said labeled binding partner is protein G and/or protein A. 
     
     
         15 . The method of  claim 12  wherein said analyte of interest comprises an anti-FMDV antibody. 
     
     
         16 . The method of  claim 15  wherein said anti-FMDV antibody is specific for a nonstructural FMDV protein. 
     
     
         17 . The method of  claim 14  wherein said anti-FMDV antibody is specific for said nonstructural FMDV protein comprising at least one of 2C, 3A, 3ABC or 3D. 
     
     
         18 . The method of  claim 12  wherein said membrane strip comprises an application point comprising a filter pad containing at least one of said labeled binding partner and said labeled control reagent. 
     
     
         19 - 50 . (canceled)