Composition, method and kit for pathology

Abstract

Disclosed herein are embodiments of compositions, methods and kits that describe ancillary reagent constructs for simultaneous immunological and morphological investigation of pathology specimens. The kits include reagents, possibly organized in panels, which give the pathologist additional confidence in making a diagnosis based on the morphology of a cell. Reagents can be used or selected to identify specific antigens or a class of antigens. The reagents further comprise detection agents, which are visible under standard microscopy in the presence of standard stains. The reagents, used on their own or as part of a kit, make possible immunological analysis simultaneously with pathology performed in the presence of standard stains.

I claim: 
     
         1 . An ancillary reagent construct comprising:
 A detection agent;   A linker; and   One or more antibody selected to bind an antigen present on a cell with a particular biological state.   
     
     
         2 . The ancillary reagent construct of  claim 1  where the detection agent is a particle ranging from 0.5-5 microns and made from a material insoluble in the solvents used in standard stains. 
     
     
         3 . The ancillary reagent construct of  claim 2  where the detection agent is selected from the group comprising a 1 micron Dynabead paramagnetic microparticle, a 2.8 micron Dynabead paramagnetic particle, a 0.8 Solulink, Nanolink™ magnetic bead cluster. 
     
     
         4 . The ancillary reagent construct of  claim 1  where the linker is produced from a biotin/streptavidin system. 
     
     
         5 . The ancillary reagent construct of  claim 1  where the linker contains a spacer to physically separate the detection agent from the antibody. 
     
     
         6 . The ancillary reagent construct of  claim 1  where the antibody is an antibody against an antigen selected from a group comprising: HBME-1, CEA, CD-3, CD-19, CD-20, kappa light chain, lambda light chain, CD 10, CD 34, MPO, CD 14, LCA or EPCAM. 
     
     
         7 . The ancillary reagent construct of  claim 1  where the detection agent is conjugated to more than one antibody selected from the group comprising HBME-1, CEA, CD-3, CD-19, CD-20, kappa light chain, lambda light chain, CD 10, CD 34, MPO, CD 14, LCA or EPCAM. 
     
     
         8 . A method for conducting simultaneous immunological and morphological investigation comprising the steps of:
 isolating a pathology specimen from a patient;   selecting an appropriate ancillary reagent construct with an antibody to an antigen expressed on a cell associated with the biological state of interest;   counting the number of cells in the specimen;   administering an appropriate amount of the ancillary reagent construct;   administering an appropriate standard stain;   identifying cells with morphology consistent with the biological state of interest; and   confirming the absence or presence of detection agents proximal to the cells with morphology consistent with the biological state of interest.   
     
     
         9 . The method of  claim 8  where:
 the patient specimen is whole blood; 
 the appropriate ancillary reagent constructs are a panel of ancillary reagent constructs conjugated to antibodies against B cell antigens and a panel of ancillary reagent constructs conjugated to antibodies against kappa and lambda light chains; and 
 the biological state is lymphocytosis. 
 
     
     
         10 . The method of  claim 9  where the B cell panel comprises ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising CD 19, CD 20, CD 3, CD24, CD72 and CD73. 
     
     
         11 . The method of  claim 8  where:
 the patient specimen is a body fluid; 
 the appropriate ancillary reagent constructs are a panel of ancillary reagent constructs are a neoplastic panel of ancillary reagent constructs; 
 and a reactive panel of ancillary reagent constructs; and 
 the biological state is cancer. 
 
     
     
         12 . The method of  claim 11  where the neoplastic panel comprises ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising: EPCAM, CEA, CD15, Berep4, ecad, and EMA. 
     
     
         13 . The method of  claim 11  where the neoplastic panel is a single ancillary reagent construct conjugated to antibodies to both EPCAM and CEA. 
     
     
         14 . The method of  claim 13  where the ratio of EPCAM to CEA on the ancillary reagent construct is substantially similar to 5:2. 
     
     
         15 . A kit for conducting simultaneous immunological and morphological analysis comprising at least one panel of ancillary reagent constructs conjugated to antibodies selected to bind antigens associated with a biological state of interest. 
     
     
         16 . A kit for differentiating neoplastic from reactive lymphocytosis comprising:
 A first panel of ancillary reagent constructs comprising reagents selected to bind B cell antigens and T cell antigens; and   A second panel of ancillary reagent constructs comprising reagents selected to bind lambda and kappa light chains.   
     
     
         17 . The kit of  claim 16  where the first panel is comprised of ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising CD 19, CD 20, CD 3, CD24, CD72 and CD73. 
     
     
         18 . The kit of  claim 16  where the second panel is comprised of ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising: lambda light chain and kappa light chain. 
     
     
         19 . A kit for identifying neoplastic cells in body fluids comprising:
 a neoplastic panel of ancillary reagent constructs; and   a reactive panel of ancillary reagent constructs.   
     
     
         20 . The kit of  claim 19  where the neoplastic panel is comprised of ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising: EPCAM, CEA, CD15, Berep4, ecad, and EMA. 
     
     
         21 . The kit of  claim 19  where the reactive panel is comprised of ancillary reagent constructs conjugated to antibodies to antigens selected from the group comprising: HBME-1 and LCA.