Immuno-pcr method

An immuno polymerase chain reaction method to detect a plurality of biological molecules in a sample wherein said method comprises the steps of i) providing a preparation comprising;

• a) an assay sample to be tested;
• and
• b) a plurality of ligands specific for at least one biological molecule in said assay sample wherein each ligand is coupled to an oligonucleotide;
• ii) incubating said preparation under conditions which allow the binding of said ligands to said biological molecules to form a complex;
• iii) adding to the first preparation a second preparation comprising;
• a) a plurality of single stranded nucleic acid templates wherein said templates comprise a nucleic acid sequence which is modified by addition, deletion or substitution of at least one nucleic acid base wherein said modifications provide a plurality of templates which vary from one another such that the annealing temperatures of said templates to at least one detectable probe molecule is varied and which at least part of each of said templates is adapted to anneal to the oligonucleotide of (b);
• and
• b) at least one oligonucleotide primer and polymerase chain reaction reagents;
• iv) conducting -a polymerase chain reaction to amplify said template;
• and
• v) conducting melt curve analysis during said reaction to determine the presence of said biological molecule.