T-Cell Receptor Gene Rearrangement Measurable/Minimal Residual Disease Assessment by Next-Generation Sequencing

T cells undergo somatic rearrangement of the T-cell receptor (TCR) genes, resulting in a high diversity of antigen binding capability. TCR is a heterodimeric protein consisting of highly variable α and β chains (αβ T-cells), or γ and δ chains (γδ T-cells). TCR gene rearrangements occur early in T-cell ontogeny and are unique to each T-cell. These rearrangements can therefore be used as a marker of a common origin, or clonality, among a T-cell population, and in the evaluation of measurable/minimal residual disease in lymphoid hematopoietic disorders. The TCR-β (TRB) gene locus is on chromosome 7(7q34) and includes 65 Vβ (variable) gene segments followed by two separate clusters of gene segments; each cluster includes a Dβ (diversity) region, a Jβ (joining) region containing several Jβ gene segments, and a Cβ (constant) region spread over 685 kilobases. The TCR-γ (TRG) gene locus is also on chromosome 7 (7q14) and includes 14 Vγ, 5 Jγ gene, and 2 Cγ gene segments spread over 200 kilobases. The Invivoscribe LymphoTrack assays are targeted next generation sequencing (NGS)-based methods that enable quantitative identification of T-cell populations using a single multiplex master mix of primers targeting the conserved V(D)J regions of the T-cell receptor γ and β (TRG and TRB) genes. The analytical sensitivity by our validation studies is one T-cell equivalent in a background of 100,000 nucleated cells with a DNA input of at least 640ng per replicate.

Quantitative detection of T cell receptor Gamma (TRG) and/or Beta (TRB) gene rearrangements in minimal residual disease specimens.