Selective determination of nucleotides in viable somatic and microbial cells
OwnerSeppo E. Kolehmainen, Veikko Tarkkaneninventors
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Abstract
A method is disclosed for measuring the concentration of nucleotides and the number of viable cells in a biological sample containing somatic and/or microbial cells by treatment of a sample of cells with surface active agents to selectively release the nucleotides without rupture of the cell membrane or walls, and measuring by means of a bioluminescent assay technique the concentration of nucleotides selectively released and thereby determining the number of viable cells in the sample.
Claims18
A method for measuring the concentration of ATP in microbial cells and the member of viable microbial cells contained in a biological sample composed of a mixture of somatic and microbial cells, which comprises contacting said sample with a non-ionic surface active agent selected from the group consisting of ethoxylated alkyl phenols and fatty acid poly glycol ethers under conditions to selectively release ATP from essentially only the somatic cell without rupturing the cell membranes of the somatic cells, removing the somatic ATP thereby released, further contacting the sample with an ionic surface active agent selected from the group consisting of ethoxylated amines, ethoxylated diamines, ethoxylated amides, polyethylene glycol esthers of fatty acids, quaternary ammonium salts, and mixtures thereof under conditions to selectively release ATP from the microbial cells without rupture of the cell membranes and cell walls of the microbial cells, and thereafter measuring by means of a bioluminsecent assay technique the concentration of ATP selectively released from the microbial cells and thereby determining the number of viable microbial cells in the sample.
A method for measuring the concentration of ATP in microbial cells and the number of viable microbial cells contained in a biological sample composed of a mixture of somatic and microbial cells, which comprises contacting said sample with a nonionic surface active agent of the formula:
- ##STR7## wherein R is an alkyl group having from 6 to 15 carbon atoms and x is from 2 to 20, under conditions to selectively release ATP from essentially only the somatic cells without rupturing the cell membranes of the somatic cells, removing the somatic ATP thereby released, further contacting the sample with a mixture of ionic surface active agents comprising a mixture of ethoxylated amine and a quaternary ammonium salt of an ethoxylated amine under conditions to selectively release ATP from the microbial cells without rupture of the cell membranes and cell walls of the microbial cells, and thereafter measuring by means of a bioluminescent assay technique the concentration of ATP selectively released from the microbial cells and thereby determining the number of viable microbial cells in the sample.
A method of measuring the concentration of ATP in microbial cells and the number of viable microbial cells contained in a biological sample containing predominantly microbial cells, comprising contacting the sample with a mixture of ionic surface active agents comprising an ethoxylated amine and a quaternary ammonium salt of an ethoxylated amine under conditions to selectively release ATP from the microbial cells without rupture of the cell membranes and cell walls of the microbial cells, and thereafter measuring by means of a firefly bioluminescent assay the concentration of ATP selectively released from the microbial cells and thereby determining the number of viable microbial cells in the sample.
Dependencies are inferred from the claim text.