Membrane assay method
The present invention provides an assay method for a cell-containing body sample, the method comprising treating the sample under conditions whereby to cause cell lysis, preferably by means of a detergent; and subjecting the thus-generated lysed sample to conditions causing the cleavage of nucleic acid molecules. The invention additionally provides the use of nucleic acid cleavage conditions in enhancing a membrane assay, a device for carrying out such an assay, and a kit for use in the assay.
1. An automated membrane assay method for at least one non-nucleic acid component of a whole blood sample, said method comprising:
treating said sample under conditions whereby to cause cell lysis thereby breaking up substantially all of the cellular membranes and nuclear membranes,
subjecting the thus-generated lysed sample to conditions causing cleavage of nucleic acid molecules, and
flowing the lysed sample through a membrane of a pore size no greater than 10 μm having immobilized thereon a specific binding ligand for the component of the sample and assaying that part of the sample retained on or by the membrane for the component of said sample, wherein said cleavage of nucleic acid molecules is provided by radiative conditions, chemical conditions and/or biological conditions.
2. The method as claimed in claim 1 wherein said biological conditions are provided by a biological reagent comprising a nuclease selected from micrococcal nuclease, S1 Nuclease, Mung Bean Nuclease, DNase I, Nuclease BAL 31 and Benzonase™.
3. The method as claimed in claim 2 wherein said nuclease is provided in dried form.
4. The method as claimed in claim 1 wherein cell lysis is detergent lysis, hypotonic lysis, hypertonic lysis or chaotropic lysis.
5. The method as claimed in claim 4 wherein said lysis is provided by a detergent selected from cationic surfactants, anionic surfactants, zwitterionic surfactants and non-ionic surfactants.
6. A method of claim 1 , wherein said nucleic acid cleavage conditions comprise:
(a) reduced membrane blockage;
(b) reduced assay time; or
(c) increased processible sample volume.
7. The method as claimed in claim 1 , wherein said one non-nucleic acid component is selected from the group consisting of small molecules, amino acids, co-factors, vitamins, oligo-peptides, poly-peptides, antibodies, proteins, protein complexes, peptide hormones, non-peptide hormones and acute-phase markers.
8. The method as claimed in claim 1 , wherein said non-nucleic acid component is selected from the group consisting of C-reactive protein (CRP), cobalamin, transcobalamin, homocysteine, folate, folate receptors and combinations thereof.
9. The method as claimed in claim 8 , wherein said non-nucleic acid component is CRP.
10. A membrane assay method for a non-nucleic acid component of a whole blood sample, said method comprising:
treating said sample under conditions whereby to cause cell lysis thereby breaking up substantially all of the cellular membranes and nuclear membranes,
subjecting the thus-generated lysed sample to enzymatic conditions causing cleavage of nucleic acid molecules, and
flowing the lysed sample through a membrane of a pore size no greater than 10 μm having immobilized thereon a specific binding ligand for the component of the sample and assaying that part of the sample retained on or by the membrane for the component of said sample.
11. The method as claimed in claim 10 wherein said enzymatic conditions are provided by a biological reagent comprising a nuclease selected from micrococcal nuclease, S1 Nuclease, Mung Bean Nuclease, DNase I, Nuclease BAL 31 and Benzonase™.
12. The method as claimed in claim 11 wherein said nuclease is provided in dried form.
13. The method as claimed in claim 10 wherein cell lysis is detergent lysis, hypotonic lysis, hypertonic lysis or chaotropic lysis.
14. The method as claimed in claim 10 wherein said lysis is provided by a detergent selected from cationic surfactants, anionic surfactants, zwitterionic surfactants and non-ionic surfactants.
15. The method as claimed in claim 10 , wherein said non-nucleic acid component is selected from the group consisting of C-reactive protein (CRP), cobalamin, transcobalamin, homocysteine, folate, folate receptors and combinations thereof.