Early detection of pathogens in blood

Abstract

The present invention is a method of extracting infectious pathogens from a volume of blood including the steps of creating a fibrin aggregate confining the pathogens and introducing a fibrin lysis reagent to expose the pathogens for analysis. The fibrin lysis reagent is preferably composed of plasminogen and streptokinase frozen in coincident relation until the fibrin lysis reagent is needed whereby streptokinase enzymatically reacts with plasminogen to form plasmin upon thawing. The plasminogen is suspended in an aqueous salt solution prior to freezing including NaCl and Na 3 PO 4 .

1 . A method of concentrating, detecting, and extracting particles from a whole blood sample, the method comprising:
 exposing the blood sample to an enzyme-detergent combination; and   analyzing the exposed blood sample for the presence of particles.   
     
     
         2 . The method of  claim 1  wherein the enzyme-detergent combination comprises plasminogen and streptokinase. 
     
     
         3 . The method of  claim 2  further comprising the steps of:
 maintaining the plasminogen and streptokinase in a frozen state in coincident relation until exposure of the blood sample to the enzyme-detergent combination; and 
 reacting streptokinase with plasminogen upon thawing whereby plasmin is formed. 
 
     
     
         4 . The method of  claim 3  wherein the plasminogen is suspended in an aqueous salt solution prior to freezing. 
     
     
         5 . The method of  claim 4  wherein the aqueous salt solution comprises NaCl. 
     
     
         6 . The method of  claim 4  wherein the aqueous salt solution comprises Na 3 PO 4 . 
     
     
         7 . The method of  claim 1  wherein the particles are selected from the group consisting of prions, toxins, metabolic markers, cancerous matter, disease state markers, bacteria, virus, and fungi. 
     
     
         8 . The method of  claim 1  wherein the particles are DNA molecules and further comprising the step of replicating the particles through PCR. 
     
     
         9 . The method of  claim 1  further comprising the step of exposing the blood sample to a DNase. 
     
     
         10 . The method of  claim 1  further comprising the step of exposing the blood sample to an endonuclease. 
     
     
         11 . The method of  claim 2  wherein the plasminogen and streptokinase are in a dried state. 
     
     
         12 . The method of  claim 11  wherein the plasminogen and streptokinase are mixed and distributed in disposable test containers. 
     
     
         13 . The method of  claim 11  wherein the plasminogen is combined with one or more enzymes selected from the group consisting of Phospholipase A 2 , DNase, Endonuclease, and Lipase. 
     
     
         14 . The method of  claim 13  wherein the enzyme-detergent combination is suspended then dried in pellets of trehalose buffer and packaged as a dry reagent. 
     
     
         15 . The method of  claim 11  wherein the streptokinase is suspended then dried in pellets of trehalose buffer and packaged into tubes as a dry reagent. 
     
     
         16 . The method of  claim 11  further comprising:
 resuspending the plasminogen and streptokinase in a buffer solution; 
 adding the buffer solution containing plasminogen and streptokinase to the volume of blood; and 
 incubating the blood sampler at room temperature. 
 
     
     
         17 . The method of  claim 16 , wherein the enzymes of the enzyme-detergent combination are in a dried state and comprise 1,500-4,500 KU Phospholipase A 2 , 5,000-10,000 U Streptokinase, 2-10 U Plasminogen, 200-3,650 U DNase, 200-4,000 U Endonuclease, and 10,000-100,000 U Lipase. 
     
     
         18 . The method of  claim 16  further comprising:
 centrifuging the buffer solution to form a supernatant and a pellet; 
 decanting the supernatant from the centrifuged buffer solution; and 
 washing the pellet. 
 
     
     
         19 . The method of  claim 18  wherein the buffer solution is centrifuged for approximately 20 minutes at 5,000-5,500×g at a temperature of 10-20° C.