Blood typing
A blood testing method for use in the detection of a disease, wherein at least one characteristic antibody or complement factor is bound to a subject's red blood cells, comprises providing a microarray wherein a plurality of binding agents therefor are immobilized on a substrate at discrete pre-defined positions; and contacting a blood sample therewith. The presence of bound red blood cells is then detected.
1. A blood testing method to confirm the detection of a disease in a subject, wherein the disease is characterised by the presence of at least one characteristic antibody/complement factor bound to the surface of the red blood cells of the subject, comprising the steps of:
providing a microarray wherein a plurality of binding agents which are capable of binding specifically to different characteristic antibodies or compliment factors on the red blood cells are immobilised on a substrate at discrete predefined positions;
contacting a blood sample from the subject with said microarray;
substantially removing any unbound red blood cells from at least an area of said substrate on which said binding agents are immobilised; and
detecting the presence or red blood cells bound to the microarray through said characteristic antibodies or complement factors, wherein the presence of any red blood cells bound to the microarray indicates the presence of said characteristic antibody/complement factor bound to the red blood cells of the subject so as to confirm detection of said disease.
2. A method according to claim 1 wherein the binding agent is a monoclonal antibody.
3. A method according to claim 1 wherein the binding agent is polyclonal antibody.
4. A method according to claim 1 wherein the binding agent is a chimeric antibody.
5. A method according to claim 1 wherein the binding agent is a single chain antibody.
6. A method according to claim 1 wherein the binding agent is selected from monoclonal anti-IgG 1 , monoclonal anti-IgG 3 and monoclonal anti-C3.
7. A method according to claim 3 wherein polyclonal anti-IgG binding agent is included.
8. A method according to claim 1 wherein at least two different binding agents are immobilised in discrete areas of the substrate.
9. A method according to claim 8 wherein each binding agent is provided in a number of different dilutions.
10. A method according to claim 9 wherein each binding agent is repeated a number of times at a given dilution.
11. A method according to claim 1 wherein the substrate is made of glass, gold silicon, silicon oxide, metals and metal oxides; either bare or functionalised with functional polymer.
12. A method according to claim 11 wherein the substrate is a gold-coated substrate.
13. A method according to claim 12 wherein the gold is functionalised such that the binding agents are capable of being immobilised thereon.
14. A method according to claim 13 wherein the functionalisation is such that the distance between the gold surface and a bound red blood cell can be controlled.
15. A method according to claim 1 wherein the microarray is formed on a planar or spheroid surface.
16. A method according to claim 1 wherein the substrate is a rigid or semi-rigid support selected from the group consisting of membranes, filter chips, slides, wafers, fibers, magnetic or non-magnetic beads, gels, tubing, plates, polymers, microparticles and capillaries.
17. A method according to claim 1 wherein the binding agent is immobilised in spots less than 1 mm in diameter.
18. A method according to claim 1 wherein the substrate comprises a plurality of separate arrays on the surface of the substrate, arranged in a manner to allow separate samples to be contacted with each array in such a way that the samples do not mix.
19. A method according to claim 1 wherein areas of the substrate not provided with binding agent are treated with blocking agents in order to minimise any non-specific binding.
20. A method according to claim 1 wherein bound red blood cells are detected by secondary labelling detection.