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Unit Code 87959:
T-Cell Receptor Excision Circles (TREC) Analysis for Immune Reconstitution

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Method Description

This assay involves isolation of peripheral blood mononuclear

cells (PBMCs) from whole blood. For adult patients, the PBMCs

are further processed to enrich for CD3 T cells, while the PBMCs

are used directly for pediatric samples. The appropriate numbers

of PBMCs are treated with proteinase K to prepare cell lysates.

The resulting cell lysates containing genomic DNA are used for

amplification in the real-time quantitative PCR assay. Each sample

will be amplified using a probe specific for the TCR delta-deletion

TREC signal joint and a probe for housekeeping gene, albumin.

There is 1 copy of TREC per T cell, while there are 2 copies of

albumin in every cell. The albumin counts are used to normalize

the cell counts in the reaction and calculation of TREC copies.

The TREC and albumin counts obtained are calculated as copies

per million cells using a standard curve for both TREC and albumin.

(Douek DC, Vescio RA, Betts MR, et al:  Assessment of thymic

output in adults after hematopoietic stem cell transplantation and

prediction of T cell reconstitution. Lancet 2000;355:1875-1881;

Douek DC, Hill B:  Personal Communication, 2005)

Performing Laboratory Location

Rochester

Key