IMAGES

Schematic representation of the two techniques used for the TREC:T-cell ratio measurements.

Schematic representation of the two techniques used for the TREC:T-cell ratio measurements.

The novel QPCR mono assay (1) using DNA purified either from PBMCs isolated by a Ficoll-Hypaque density gradient centrifugation or from dried blood spot samples, and the usual assay combining QPCR measurement for TREC measurement (2) and FACS analysis for the T-cell count (3). The figure demonstrates how the approximation of the VD-J to T-cell count is derived with the novel QPCR mono assay (1). From either whole blood samples or dried blood spots samples, the total cellular DNA is quantified by using the count for ALB2 (albumin gene) considered as a housekeeping gene (A). Similarly, unrearranged TCR-β count is quantified according to the VD-J level; unrearranged TCR-β cells correspond to the total germline cell number (B). Therefore the unrearranged TCR-β PBMC count subtracted from the total PBMC count give us the final rearranged TCR- β count and hence the T-cell count (A – B).

You are in

Biomaterials