In Vitro Diagnosis Device and Uses Thereof

Abstract

The invention relates to an in vitro diagnosis device for the detection of at least one reaction between an erythrocytic phenotype antigen and an antibody specifically directed against said antigen in a sample of blood or one of the components thereof. The device is characterized in that it comprises: a substrate; and a hydrophobic porous membrane having a thickness of between 0.5 mm and 1.5 mm and a pore diameter of between 2 and 30 μm, said membrane comprising at least one hydrophilic reaction zone intended to receive the sample. The invention also relates to the uses of said device in immunohematology.

1 . An in-vitro diagnosis device ( 10 ), for detection of at least one reaction between an antigen of erythrocytic phenotype and an antibody directed specifically against this antigen, from a sample of blood or one of its components, characterized in that it comprises:
 a support ( 12 ), and   a hydrophobic porous membrane ( 14 ) of thickness between 0.05 mm and 1.5 mm and whereof the diameter of the pores is between 2 and 30 μm, said membrane comprising at least one hydrophilic reaction area ( 16 ) intended to receive said sample, the surface of the hydrophilic reaction area ( 16 ) being less than the surface of the hydrophobic porous membrane ( 14 ).   
     
     
         2 . The diagnosis device ( 10 ) according to  claim 1 , characterized in that the hydrophilic reaction area ( 16 ) of the porous membrane ( 14 ) has been made hydrophilic with a detergent without modification of the chemical functions of the porous substrate by prior chemical or physical treatment of the porous membrane. 
     
     
         3 . The diagnosis device ( 10 ) according to  claim 2 , characterized in that the detergent is a non-ionic surfactant. 
     
     
         4 . The diagnosis device ( 10 ) according to  claim 2  or  3 , characterized in that the detergent is used at a dose between 0.01 and 2% (weight/volume). 
     
     
         5 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the porous membrane ( 14 ) is arranged in the support ( 12 ) and in that the support ( 12 ) comprises at least one opening ( 20 ), each opening ( 20 ) being at right angle of each hydrophilic reaction area ( 16 ) of the porous membrane. 
     
     
         6 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the support ( 12 ) is a rigid plastic support. 
     
     
         7 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that it also comprises an absorbent membrane ( 18 ) arranged underneath the porous membrane ( 14 ). 
     
     
         8 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the hydrophilic reaction area ( 16 ) of the porous membrane ( 14 ) also comprises capturing agents. 
     
     
         9 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the capturing agents comprise antigens of erythrocytic group/phenotype. 
     
     
         10 . The diagnosis device ( 10 ) according to  claim 9 , characterized in that the capturing agents are red blood cells void of haemoglobin. 
     
     
         11 . The diagnosis device ( 10 ) according to any one of  claims 1  to  8 , characterized in that the capturing agents are antibodies. 
     
     
         12 . The diagnosis device ( 10 ) according to any one of  claims 1  to  8 , characterized in that the capturing agents are polycationic polymers. 
     
     
         13 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the reaction area ( 16 ) is hydrophilic over the entire thickness of the porous membrane. 
     
     
         14 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the reaction area ( 16 ) is hydrophilic at the surface of said reaction area. 
     
     
         15 . The diagnosis device ( 10 ) according to any one of the preceding claims, characterized in that the reaction area ( 16 ) comprises two hydrophilic areas ( 16 - 1 ,  16 - 2 ) with a greater degree of hydrophilisation at the centre ( 16 - 1 ) of the reaction area than at the periphery ( 16 - 2 ). 
     
     
         16 . The diagnosis device ( 10 ) according to any one of  claims 1  to  14 , characterized in that the reaction area ( 16 ) comprises two hydrophilic areas with a different degree of hydrophilisation, one at the surface, the other in the thickness. 
     
     
         17 . The diagnosis device ( 10 ) according to  claim 15  or  16 , characterized in that the reaction area ( 16 ) has been made hydrophilic with two different detergents without modification of the chemical functions of the porous substrate by prior chemical or physical treatment of the porous membrane. 
     
     
         18 . A manufacturing process of a device ( 10 ) according to any one of the preceding claims, characterized in that it comprises the following steps:
 hydrophilisation of a hydrophobic porous membrane ( 14 ) of thickness between 0.05 mm and 1.5 mm and whereof the diameter of the pores is between 2 and 30 μm, on at least one area ( 16 ) of said membrane, by means of at least one detergent,   optionally depositing of a capturing agent solution,   drying,   assembling of the porous membrane ( 14 ) and optionally an absorbent membrane ( 18 ) arranged underneath the porous membrane ( 14 ) with a support ( 12 ).   
     
     
         19 . Use of the device according to any one of  claims 1  to  17 , to identify and determine ABO blood groups, extended Rhesus phenotyping, search for irregular agglutinin, search for autoantibodies, search for cold antiglobulin and/or cross-validation, from a sample of blood or one of its components. 
     
     
         20 . A phenotyping process of erythrocytic blood groups, from a sample of red globules, for detection of the presence of two different populations of antigens, characterized in that it comprises the following steps:
 depositing a solution to hydrate the porous membrane ( 14 ) at the centre of the reaction area of a device ( 10 ) according to any one of  claims 1  to  7 , said reaction area ( 16 ) comprising two hydrophilic areas ( 16 - 1 ,  16 - 2 ) with a degree of hydrophilisation greater at the centre ( 16 - 1 ) of the reaction area than at the periphery ( 16 - 2 ), and comprising at the centre capturing agents comprising antibodies,   diluting the red globules to be phenotyped in a buffer solution,   adding this solution containing the red globules to be phenotyped at the centre of the reaction area,   incubating,   depositing a rinse solution on the reaction area.   
     
     
         21 . The phenotyping process of erythrocytic blood groups, from a sample of red globules, characterized in that it comprises the following steps:
 depositing a solution to hydrate the porous membrane ( 14 ) at the level of the reaction area ( 16 ) of a device ( 10 ) according to any one of  claims 1  to  7 , said reaction area ( 16 ) comprising a single hydrophilic area and comprising capturing agents comprising antibodies,   optionally diluting the red globules to be phenotyped in a buffer solution,   adding this solution containing the red globules to be phenotyped at the centre of the reaction area,   incubating,   depositing a rinse solution on the reaction area.   
     
     
         22 . The process for detection of multivalent antibodies present in the blood, from a sample of plasma, serum or total blood, characterized in that it comprises the following steps:
 depositing the sample to be tested on the reaction area ( 16 ) of a device ( 10 ) according to any one of  claims 1  to  7 , said reaction area ( 16 ) comprising capturing agents comprising antigens,   adding red blood cell tests of known phenotype, comprising the same antigens as the capturing agents,   having the mixture pass through the hydrophilic area,   depositing a rinse solution on the reaction area.   
     
     
         23 . The process for detection of anti-erythrocytic antibodies present in the blood, cross validation or search for autoantibodies or cold agglutinin from a sample of plasma, serum or total blood, characterized in that it comprises the following steps:
 incubating the sample to be tested with a buffer, red blood cell tests of known phenotype,   adding to this mixture an agent capable of aggregating the red globules,   depositing the mixture on the reaction area ( 16 ) of a device ( 10 ) according to any one of  claims 1  to  7 ,   depositing a solution containing an agent capable of aggregating the red globules on the reaction area,   depositing a Coombs, human antiglobulin or anti-complement reagent on the reaction area, and   depositing a rinse solution on the reaction area.