Generic Multiplex Digital PCR for Accurate Quantification of T Cells in Copy Number Stable and Unstable DNA Samples.
Methods Mol Biol
; 2453: 191-208, 2022.
Article
em En
| MEDLINE
| ID: mdl-35622328
ABSTRACT
An accurate T cell quantification is prognostically and therapeutically relevant in various clinical applications, including oncology care and research. In this chapter, we describe how T cell quantifications can be obtained from bulk DNA samples with a multiplex digital PCR experiment. The experimental setup includes the concurrent quantification of three different DNA targets within one reaction a unique T cell DNA marker, a regional corrector, and a reference DNA marker. The T cell marker is biallelically absent in T cells due to VDJ rearrangements, while the reference is diploid in all cells. The so-called regional corrector allows to correct for possible copy number alterations at the T cell marker locus in cancer cells. By mathematically integrating the measurements of all three markers, T cells can be accurately quantified in both copy number stable and unstable DNA samples.
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Base de dados:
MEDLINE
Assunto principal:
Variações do Número de Cópias de DNA
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Reação em Cadeia da Polimerase Multiplex
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article