Method for measuring a change in an individual's immunorepertoire

A method of calculating the change in the immune repertoire of a subject, comprising the steps of:

• collecting at least two sample populations of white blood cells from a subject;
• separately amplifying polynucleotides from each population of white blood cells in a reaction mix comprising target-specific nested primers to produce a set of first amplicons, at least a portion of the target-specific nested primers comprising additional nucleotides which, during amplification, serve as a template for incorporating into the first amplicons a binding site for at least one common primer;
• separately transferring a portion of each of the first reaction mixes containing the first amplicons to second reaction mixes comprising at least one common primer;
• for each population, amplifying, using the at least one common primer, the first amplicons to produce a set of second amplicons;
• sequencing each of the second amplicons to identify the frequency of specific CDR3 regions present in the sample;
• normalizing the CDR3 frequency data;
• comparing the normalized CDR3 frequency data of the first sample and the second sample to determine the absolute difference between the frequency of each CDR3 sequence;
• and
• determining the CDR3 sequences, common to the first sample and the second sample, with the largest frequency changes.