Quick method for detecting enyzmes and microorganisms

Abstract

The present invention concerns an in vitro method for detecting an enzyme or a microorganism in a biological sample, comprising the steps of: a1) concentrating the microorganisms present in the biological sample, optionally after a a0) culture step of the microorganisms; b1) placing in suspension the microorganisms concentrated at step a1) in a solution containing at least one chromogenic or fluorogenic substrate capable of releasing a chromophore or fluorophore after hydrolysis by the enzyme to be detected; c1) detecting potential release of the chromophore or fluorophore obtained at step b1); the release of the chromophore or fluorophore detected at step c1) indicating the presence of the enzyme to be detected.

1 - 12 . (canceled) 
     
     
         13 . A method for the in vitro preparation of a blood culture sample containing microorganisms, comprising the steps of:
 A) lysing or agglutinating the red blood cells present in the blood culture sample, without lysing the microorganisms present in the blood culture sample,   B) separating the microorganisms present in the blood culture sample from the red blood cells lysed or agglutinated at step A), and   C) washing the microorganisms of the blood culture sample separated at step B).   
     
     
         14 . The method according to  claim 13 , wherein the microorganisms are microorganisms resistant to 3rd generation cephalosporins. 
     
     
         15 . The method according to  claim 13 , wherein the lysis step A) comprises placing the blood culture sample in contact with a lysis buffer of red blood cells for a suitable time at a suitable temperature to allow lysis of the red blood cells. 
     
     
         16 . The method according to  claim 13 , wherein the agglutination step A) is implemented by placing the blood culture in contact with at least one agglutinating agent. 
     
     
         17 . The method according to  claim 16 , wherein the agglutinating agent is polyethylene glycol (PEG). 
     
     
         18 . The method according to  claim 13 , wherein the separation step B) is implemented by centrifugation, filtration or removing the aqueous phase, preferably after decantation. 
     
     
         19 . The method according to  claim 18 , wherein the separation step B) is implemented by centrifugation. 
     
     
         20 . The method according to  claim 13 , wherein the washing step C) is performed by re-suspending the separated microorganisms in a washing buffer, incubating for a suitable time, at a suitable temperature and further separating the microorganisms.