Method of diagnosis of foot and mouth disease and the diagnostic kit

Abstract

The present invention provides a method for diagnosing foot-and-mouth disease virus infection, comprising the steps of applying a predetermined amount of a test sample to a loading region of a strip; coupling a detection reagent including a given labeling reagent to an analyte of interest in the test sample to form a complex therebetween; developing the complex onto a wicking membrane; and observing changes in appearance of a reactivity zone having at least more than one immobilized phase selected from antigen, antibody or hapten on the predetermined region of the wicking membrane, derived from FMDV or obtainable from FMDV through an immunological reaction to determine the presence or absence of foot-and-mouth disease virus infection. It also provides a kit for diagnosing foot-and-mouth disease virus infection comprising a strip 1 including a reactivity zone 13 containing at least more than one immobilized phase selected from antigen, antibody or hapten thereon, derived from FMDV or obtainable from FMDV through an immunological reaction, and a control zone 14 for confining normal operation of the kit, provided on a predetermined region of a wicking membrane 9 ; and a housing 20 protecting the strip 1 from a variety of contaminants, and including at least a test sample application port 2 and an indicia window 4 for observing results of reaction in the reactivity zone 13 and the control zone 14 on the strip.

1 . A method for diagnosing foot-and-mouth disease virus infection, comprising the steps of: 
 (1) applying a predetermined amount of a test sample to a loading region of a strip; (2) coupling a detection reagent including a given labeling reagent to an analyte of interest in the test sample to form a complex therebetween; (3) developing the complex onto a wicking membrane; and (4) observing changes in appearance of a reactivity zone having at least more than one immobilized phase selected from antigen, antibody or hapten on the predetermined region of the wicking membrane, derived from FMDV or obtainable from FMDV through an immunological reaction to determine the presence or absence of foot-and-mouth disease virus infection.    
     
     
         2 . The method as set forth in  claim 1 , wherein the method is a sandwich method or a competition method.  
     
     
         3 . The method as set forth in  claim 1 , wherein the immobilized phase contains at least more than one protein selected from FMDV-derived non-structural proteins and/or structural proteins.  
     
     
         4 . The method as set forth in  claim 1  or  3 , wherein the immobilized phase is the structural proteins VP1-VP4 polypeptide or an inactivated FMDV virus disrupted material.  
     
     
         5 . The method as set forth in  claim 1  or  3 , wherein the immobilized phase contains at least more than one polypeptide selected from the group consisting of non-structural proteins leader peptide (Lb), 2B, 2C, 3D, 3A, 3AB and 3ABC.  
     
     
         6 . The method as set forth in  claim 1 , wherein the reactivity zone includes a first reactivity zone containing at least one selected from the structural protein and 3D protein, these being capable of vaccine production; and a second reactivity zone containing at least one selected from the non-structural protein, except for 3D.  
     
     
         7 . The method as set forth in  claim 1 , wherein the detection reagent is provided at any site on the strip.  
     
     
         8 . The method as set forth in  claim 7 , wherein the detection reagent includes at least more than one material selected from labeled antigens, antibodies or haptens.  
     
     
         9 . The method as set forth in  claim 7 , wherein the detection reagent includes at least more than one material selected from the group of antibodies capable of binding to labeled protein G, protein A, protein G/A, or species-specific IgG and IgM.  
     
     
         10 . The method as set forth in  claim 1 , wherein the test sample is blood, serum, plasma, urine, tears, saliva or milk.  
     
     
         11 . A kit for diagnosing foot-and-mouth disease virus infection comprising: 
 a strip including a reactivity zone containing at least more than one immobilized phase selected from antigen, antibody or hapten thereon, derived from FMDV or obtainable from FMDV through an immunological reaction, and a control zone for confirming normal operation of the kit, provided on a predetermined region of a wicking membrane; and    a housing receiving the strip, and including at least a test sample application port and an indicia window for observing results of reaction in the reactivity zone and the control zone on the strip.    
     
     
         12 . The kit as set forth in  claim 11 , wherein the immobilized phase contains at least more than one protein selected from FMDV-derived non-structural proteins and/or structural proteins.  
     
     
         13 . The kit as set forth in  claim 11  or  12 , wherein the immobilized phase is the structural protein VP1-VP4 polypeptide or an inactivated FMDV virus disrupted material.  
     
     
         14 . The kit as set forth in  claim 8 , wherein the immobilized phase contains at least more than one polypeptide selected from the group consisting of non-structural proteins leader peptide (Lb), 2B, 2C, 3D, 3A, 3AB and 3ABC.  
     
     
         15 . The kit as set forth in  claim 11 , wherein the reactivity zone includes a first reactivity zone containing at least one selected from the structural protein and 3D protein which can be used in vaccine production; and a second reactivity zone containing at least one selected from the non-structural proteins, except for 3D.  
     
     
         16 . The kit as set forth in  claim 11 , wherein the detection reagent is provided at any site on the strip.  
     
     
         17 . The kit as set forth in  claim 16 , wherein the detection reagent includes at least more than one material selected from a group of antibodies capable of binding to labeled protein G, protein A, protein G/A, or species-specific IgG and IgM.  
     
     
         18 . The kit as set forth in  claim 11 , wherein the strip has a configuration in which a reservoir pad, a filter pad in contact with one end of the reservoir pad for filtering the test sample, a wicking membrane in contact with one end of the filter pad for moving the filtered sample by capillary action, and an absorbent pad in contact with one end of the wicking membrane are connected sequentially on a base member.  
     
     
         19 . The kit as set forth in  claim 18 , wherein the detection reagent is provided at any site on the strip.  
     
     
         20 . The kit as set forth in  claim 18 , wherein the base member is made of plastic or glass material.  
     
     
         21 . The kit as set forth in  claim 18 , wherein the detection reagent contains a predetermined label such that an analyte of interest can be confirmed by naked eye or other instrumentation from the outside.  
     
     
         22 . The kit as set forth in  claim 18 , wherein the detection reagent contains at least one label selected from the group consisting of catalyst, enzyme, substrate for enzyme, fluorescent compound, chemoluminescent compound, radioactive element, metal sol, non-metal sol, dye sol, color indicator, color matter contained in liposome, latex particle, and immunochemical label.  
     
     
         23 . The kit as set forth in  claim 11 , wherein the analyte in the test sample is antigen, antibody or hapten and any combinations thereof.  
     
     
         24 . The kit as set forth in  claim 23 , wherein the antibody is monoclonal or polyclonal antibody.  
     
     
         25 . The kit as set forth in  claim 18 , wherein the strip further comprises another filter pad between the reservoir pad and the filter pad.  
     
     
         26 . The kit as set forth in  claim 11 , wherein material for the wicking membrane is at least more than one material selected from the group consisting of nylon, polyester, cellulose, polysulfone, polyvinylidenedifluoride, cellulose acetate, polyurethane, glass fiber and nitrocellulose.  
     
     
         27 . The kit as set forth in  claim 11 , wherein the immobilized phase constituting a control zone is one selected from avidin, biotin, FITC, anti-FITC antibody, mouse immunoglobulin and anti-mouse immunoglobulin antibody.  
     
     
         28 . The kit as set forth in  claim 11 , wherein the control reagent is selected from labeled protein, antigen and antibody which specifically bind to the immobilized phase constituting the control zone on the wicking membrane.