Kits and Devices for Performing Methods of Detecting Viability-Associated Molecules

Abstract

Kits and devices for performing a method of detecting a molecule associated with viability of one or more cells or organisms in a sample are provided. The method comprises the initial step of contacting the sample with an enzyme, which enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule in the presence of the molecule associated with viability of the one or more cells or organisms. This thereby generates a novel detectable nucleic acid molecule. The next step involves detecting the presence of the molecule associated with viability of the one or more cells or organisms by detecting the novel nucleic acid molecule generated only in the presence of the molecule associated with viability of the one or more cells or organisms. A most preferred molecule associated with viability is ATP, although NAD may also be detected. A preferred enzyme for use in the methods is ligase. The kits and devices for performing the method have numerous applications, in particular in monitoring viability of cells, toxicology testing and determining whether there is contamination in a sample or on a surface.

1 .- 70 . (canceled) 
     
     
         71 . A kit for any of:
 (i) detecting a molecule associated with viability from one or more cells or organisms in a sample, and/or   (ii) screening for resistance of one or more cells or organisms to a molecule directed against said cells or organisms, and/or   (iii) screening for molecules which are capable of killing or preventing growth of one or more cells or organisms, and/or   (iv) determining the toxicity of a candidate pharmaceutical agent or agricultural agent to mammalian or plant cells comprising:
 (a) a nucleic acid molecule, 
 (b) a ligase capable of adding a chemical moiety to the nucleic acid molecule in the presence of the molecule associated with viability of the one or more cells or organisms, thereby generating a novel detectable nucleic acid molecule; and 
 (c) a further nucleic acid molecule; 
   wherein the ligase is provided in a deactivated form lacking an activator co-factor or the kit further comprises means for de-activating the ligase prior to use.   
     
     
         72 . The kit according to  claim 71  wherein the ligase comprises T4 DNA ligase, T4 RNA ligase or  E. coli  DNA ligase. 
     
     
         73 . The kit according to  claim 71  wherein the enzyme and/or nucleic acid molecule and/or further nucleic acid molecule is immobilized on a solid support. 
     
     
         74 . The kit according to  claim 71  further comprising reagents necessary for nucleic acid amplification. 
     
     
         75 . The kit according to  claim 71  which comprises a filter to separate the one or more cells or organisms to be detected from other cells. 
     
     
         76 . The kit according to  claim 71  which further comprises an enzyme for removing or exhausting from the sample any molecules associated with viability of the one or more cells or organisms which are not provided by the one or more cells or organisms. 
     
     
         77 . The kit according to  claim 71  which further comprises reagents for lysis of the cells of the one or more cells or organisms in order to release the molecule associated with viability of the one or more cells or organisms. 
     
     
         78 . The kit according to  claim 71  which further comprises one or more nucleases in order to degrade nucleic acid molecules associated with the one or more cells or organisms. 
     
     
         79 . The kit according to  claim 71  which is in the foam of a homogeneous reaction mix. 
     
     
         80 . A spray device containing the homogenous reaction mix according to  claim 79  which is for administering the homogenous reaction mix to a surface.