Nucleic acid amplification controls

Abstract

The present invention discloses positive control material for nucleic acid amplification based detection of microorganisms in biological samples. The control material comprises purified microorganism that is rendered non-infectious but is amenable to nucleic acid amplification. Also disclosed is a process for making and using the control material.

1 . A method for making a full process positive control material for detection of virus in biological samples comprising:
 a) purifying an intact virus from a source;   b) exposing the purified intact virus to an aldehyde at a temperature and for a time such that one or more surface proteins are irreversibly modified while leaving the nuclear components substantially intact thereby rendering the purified intact virus non-pathogenic and wherein nucleic acids of the purified intact virus are amenable to amplification; and   c) identifying that the purified intact virus can be used as a full process positive control by confirming absence of active virus and an ability of viral nucleic acids in the purified intact virus to be amplified.   
     
     
         2 . The method of  claim 1 , wherein the aldehyde is selected from the group consisting of paraformaldehyde, formaldehyde, acetaldehyde, propionaldehyde, n-butyraldehyde, benzaldehyde, p-nitrobenzaldehyde, p-tolualdehyde, salicylaldehyde, phenylacetaldehyde, 2-methylpentanal, 3-methylpentanal, and 4-methylpentanal. 
     
     
         3 . The method of  claim 1 , wherein the time is controlled by quenching the aldehyde to provide the exposing for the time such that one or more surface proteins are irreversibly modified while leaving the nuclear components substantially intact. 
     
     
         4 . The method of  claim 3 , wherein the quenching is carried out by glycine. 
     
     
         5 . The method of  claim 1 , further comprising storing the purified intact virus at a refrigeration temperature. 
     
     
         6 . The method of  claim 1 , further comprising suspending the purified intact virus in a liquid matrix comprising a buffer, a biological fluid, or a synthetic biological fluid. 
     
     
         7 . A non-pathogenic intact virus prepared by the method of  claim 1 , wherein the nucleic acids in the non-pathogenic intact virus are amenable to amplification. 
     
     
         8 . A non-pathogenic intact virus prepared by the method of  claim 1 , wherein the nucleic acids in the non-pathogenic intact virus are amenable to amplification after storage at a refrigeration temperature.