System for detecting bacteria in blood, blood products, and fluids of tissues

Abstract

The invention provides methods for screening for the presence of a clinically relevant amount of bacteria in donor blood or a blood product from a donor mammal, particularly blood or a blood product that will be transferred from the donor mammal to a recipient mammal. The method comprises contacting a sample of the donor blood or a blood product with a set of binding agents that comprises binding agents that specifically bind to Gram-negative bacterial antigen and/or binding agents that specifically bind to Gram-positive bacterial antigen, and determining binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-positive bacteria and/or Gram-negative bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of Gram-positive bacteria and/or Gram-negative bacteria in the donor blood or blood product. The invention further provides methods and kits for screening for the presence of a clinically relevant amount of Gram-positive bacteria, Gram-negative bacteria, or both Gram-positive and Gram-negative bacteria in a donor tissue by screening the fluid in which the donor tissue is stored.

1 . A method for screening for the presence of a clinically relevant amount of bacteria in donor blood or blood product from a donor mammal for transfer to a recipient mammal comprising contacting a sample of the donor blood or blood product with a set of binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-negative bacterial antigen and binding agents that specifically bind to a Gram-positive bacterial antigen, and determining binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of bacteria in the donor blood or blood product.  
     
     
         2 - 35 . (Cancelled)  
     
     
         36 . A method for screening for the presence of a clinically relevant amount of bacteria in donor blood or blood product from a donor mammal for transfer into a recipient mammal, comprising: contacting a sample of the donor blood or blood product with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-negative bacterial antigen and binding agents that specifically bind to a Gram-positive bacterial antigen, detecting binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of bacteria in the donor blood or blood product from the donor mammal, wherein the blood or blood product determined to have less than 1×10 6  colony forming units (CFU) per mL of bacteria is useful for transfer to the recipient mammal.  
     
     
         37 . The method of  claim 36 , wherein the donor blood or blood product is selected from the group consisting of whole blood, leukocytes, hematopoietic stem cells, platelets, red blood cells, plasma, and serum.  
     
     
         38 . The method of  claim 36 , wherein the binding agents that specifically bind to the Gram-negative bacterial antigen specifically bind to the lipopolysaccharide structure of the Gram-negative bacteria.  
     
     
         39 . The method of  claim 36 , wherein the binding agents that specifically bind to the Gram-positive bacterial antigen specifically bind to the lipoteichoic acid structure of the Gram-positive bacteria.  
     
     
         40 . The method of  claim 36 , wherein the set of binding agents is immobilized on a solid-phase support.  
     
     
         41 . A method for screening for the presence of a clinically relevant amount of Gram-positive bacteria in donor blood product from a donor mammal for transfer into a recipient mammal, comprising: contacting a sample of the donor blood or blood product with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-positive bacterial antigen, detecting binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-positive bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of Gram-positive bacteria in the donor blood or blood product, and wherein the donor blood or blood product from the donor mammal determined to have less than 1×10 6  CFU per mL of Gram-positive bacteria is useful for transfer to the recipient mammal.  
     
     
         42 . A method for screening for the presence of a clinically relevant amount of Gram-negative bacteria in donor blood product from a donor mammal for transfer to a recipient mammal, comprising: contacting a sample of the donor blood or blood product with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-negative bacterial antigen, detecting binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-negative bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of Gram-negative bacteria in the donor blood or blood product, and wherein the donor blood or blood product from the donor mammal determined to have less than 1×10 6  CFU per mL of Gram-negative bacteria is useful for transfer to the recipient mammal.  
     
     
         43 . A method for screening for the presence of a clinically relevant amount of bacteria in a donor tissue from a donor mammal for transfer to a recipient mammal, wherein the donor tissue is stored in a fluid, comprising contacting a sample of the fluid with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-negative bacterial antigen and binding agents that specifically bind to a Gram-positive bacterial antigen, detecting binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of bacteria in the donor tissue and no binding indicates the absence of a clinically relevant amount of bacteria in the donor tissue, and wherein the donor tissue from the donor mammal determined to have less than 1×10 6  CFU per mL of bacteria is useful for transfer to the recipient mammal.  
     
     
         44 . The method of  claim 43 , wherein the donor tissue determined to have an absence of a clinically relevant amount of bacteria is transferred to the second mammal.  
     
     
         45 . The method of  claim 43 , wherein the donor tissue is selected from the group consisting of lung, heart, liver, skin, kidney, pancreas, spleen, and bone marrow.  
     
     
         46 . A method for screening for the presence of a clinically relevant amount of Gram-positive bacteria in a donor tissue from a donor mammal for transfer to a recipient mammal, wherein the donor tissue is stored in a fluid, comprising contacting a sample of fluid with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-positive bacterial antigen, detecting binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-positive bacteria in the donor tissue and no binding indicates the absence of a clinically relevant amount of Gram-positive bacteria in the donor tissue, and wherein the donor tissue from the donor mammal determined to have less than 1×10 6  CFU per mL of Gram-positive bacteria is useful for transfer to the recipient mammal.  
     
     
         47 . A method for screening for the presence of a clinically relevant amount of Gram-negative bacteria in a donor tissue from a donor mammal for transfer to a recipient mammal, wherein the donor tissue is stored in a fluid, comprising contacting a sample of the fluid with a set of pan-generic binding agents, wherein the set of binding agents comprises binding agents that specifically bind to a Gram-negative bacterial antigen, detecting binding of the set of binding agents to the Gram-negative bacterial antigen in the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-negative bacteria in the donor tissue and no binding indicates the absence of a clinically relevant amount of Gram-negative bacteria in the donor tissue, and wherein the donor tissue from the donor mammal determined to have less than 1×10 6  CFU per mL of Gram-negative bacteria is useful for transfer to the recipient mammal.  
     
     
         48 . The method of any of claims  36 ,  41 ,  42 ,  43 ,  46 , and  47 , wherein the set of binding agents are detectably labeled with a reporter molecule.  
     
     
         49 . The method of  claim 48 , wherein said reporter molecule is selected from the group consisting of a molecule with enzymatic activity, a radio-labeled molecule, a fusion molecule, a fluorogenic molecule, a metal sol, a particle, a chromatic molecule, and a molecule that is specifically bound by a secondary agent.  
     
     
         50 . The method of any of claims  36 ,  41 ,  42 ,  43 ,  46 , and  47 , wherein the clinically effective amount of bacteria is greater than 1×10 5  CFU/ml of blood, blood product or tissue.  
     
     
         51 . The method of any of claims  36 ,  41 ,  42 ,  43 ,  46 , and  47 , wherein the clinically effective amount of bacteria is greater than 1×10 4  CFU/ml of blood, blood product or tissue.  
     
     
         52 . The method of any of claims  36 ,  41 ,  42 ,  43 ,  46 , and  47 , wherein the clinically effective amount of bacteria is greater than 1×10 3  CFU/ml of blood, blood product, or tissue.  
     
     
         53 . The method of  claim 49 , wherein the enzymatic molecule is selected from the group consisting of horseradish peroxidase, alkaline phosphatase, and β-galactosidase.  
     
     
         54 . The method of  claim 48 , wherein said reporter molecule is bound to the binding agent by intermolecular association.  
     
     
         55 . The method any of claims  36 ,  41 ,  42 ,  43 ,  46 , and  47  further comprising the step of transferring the blood, blood product, or tissue to a recipient mammal.