Use of Ribozymes in the Detection of Adventitious Agents

Abstract

The present invention provides a method of detecting adventitious agents in a composition comprising a microorganism by using ribozyme-expressing indicator cells, as well as indicator cells useful in such detection.

1 - 31 . (canceled) 
     
     
         32 . A method of detecting the presence of an adventitious agent in a therapeutic composition comprising a reovirus, the method comprising the steps of:
 (a) providing a population of indicator cells that expresses a ribozyme, wherein the ribozyme is capable of specifically cleaving the genome of the reovirus;   (b) contacting the indicator cells with the therapeutic composition comprising a reovirus under conditions that allow for cleavage of the genome of the reovirus by the ribozyme; and   (c) detecting the presence or absence of a pathogenic effect on the indicator cells contacted with the therapeutic composition, wherein the presence of a pathogenic effect indicates the presence of an adventitious agent in the therapeutic composition.   
     
     
         33 . The method of claim  1  wherein the indicator cells express the ribozyme from a gene that is integrated into the genome of the cells. 
     
     
         34 . The method of claim  1  wherein the ribozyme is capable of specifically cleaving the sl RNA of the reovirus. 
     
     
         35 . The method of claim  1  wherein the ribozyme is Rz-553 or Rz-984. 
     
     
         36 . The method of claim  1  wherein the reovirus is a mammalian reovirus or a human reovirus. 
     
     
         37 . The method of claim  1  wherein the reovirus is a serotype 3 reovirus. 
     
     
         38 . The method of claim  1  wherein the reovirus is a recombinant reovirus. 
     
     
         39 . The method of claim  1 , wherein the population of indicator cells comprises human cells. 
     
     
         40 . The method of claim  1 , wherein the adventitious agent is an infectious agent, other than reovirus, which is capable of infecting the indicator cell.