Molecular Assays of T-cell Receptors

For any of the following assays please fill out and submit the appropriate form.

If you are using a FHCRC project id for billing, please fill out the Internal Work Order Form.

If billing is to be submitted to the University of Washington or other external institution, fill out the External Work Order Form.

TCR Spectratyping

Spectratyping is a set of multiplex RT-PCR reactions that amplifies all T-cell receptor beta families to reveal the clonal length polymorphism in the CDR3 region. This technique is used to access clonal composition in T-cell populations. Specifically, it is often used to determine clonal expansion, contraction and deletion during immune response and reconstitution. TCR spectratyping can be provided for PBMC samples or for FACS-sorted cell subsets.

Clonal TCR analysis and tracking by real-time PCR

The unique feature of each T-cell clone is found in the recombined T cell receptor sequences. Immune Monitoring can identify the clonal TCR beta chain sequence by RT-PCR and direct sequencing of the clonal cell sample. The clonal TCRB sequence can be cloned and used for QPCR analysis.

With knowledge of the clonal TCR sequence, specific primers and a probe for quantitative real-time PCR analysis can be designed, allowing the quantitation of the clonal T cells in a cell population. We provide this service in the setting of T cell adoptive transfers in immune therapy trials of cancer patients.

TREC assay

TREC is a quantitative real-time PCR analysis of the TCR excision circle, a circular DNA fragment excised from the genome as a result of the TCR rearrangement in the thymus. This circular DNA is not capable of replication and is thereore diluted as T cells divide in the periphery. The relative quantity of the TREC DNA provides a measurement of thymic output of T cells.