METHODS OF SEQUENCING THE IMMUNE REPERTOIRE

摘要

The invention provides a non-invasive technique for the detection and quantification of the immune repertoire, in a biological sample containing a plurality of distinct cell populations. Methods are conducted using sequencing technology to detect and enumerate immune repertoire within a heterogeneous biological sample.

主权项

1. A method of determining the immune repertoire in a subject, comprising:
a. isolating a plurality of RNA from a biological sample comprising a plurality of cell types obtained from a subject, b. producing immunoglobulin chain or TCR chain cDNAs from the RNA; c. adding a homopolymeric tail, a random molecular tag and a universal sequence to the 3′ end of the cDNAs; d. amplifying the cDNAs by using a one or more immunoglobulin chain or TCR chain specific primers and a universal sequence specific primer comprising an flanking sequence specific to the sequencing platform to produce a plurality of molecular tagged immunoglobulin chain or TCR chain nucleic acids; e. sequencing the amplified immunoglobuling chain or TCR chain nucleic acids to produce a plurality of sequencing reads thereby determining the immune repertoire of the subject.