Rapid screening of T-cell receptor (TCR) variable gene usage by multiplex PCR: Application for assessment of clonal composition

Y. Akatsuka, E. G. Martin, A. Madonik, A. A. Barsoukov, John A. Hansen

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The selection of various T-cell receptor (TCR) gene families and complex rearrangements during intra-thymic differentiation provide the basis for the expression of antigen specificity by mature T cells. TCR beta variable (TCRBV) transcripts can be identified by RT-PCR, but multiple reactions are required to detect all genes of the TCRBV subfamilies. We describe here a multiplex PCR method that amplifies 46 functional genes comparing 23 TCRBV families in 5 reactions where each reaction contains 4 to 7 specific primers together with a single fluorescence-tagged TCR beta constant region primer. Between 8 and 10 distinct subtypes within each of the 23 TCRBV families can be identified by analysis of the CDR3 length. Multiplex PCR products isolated from agarose gels can be subjected to direct sequencing for confirmation and definitive clonotyping if necessary. The data illustrated here show that the multiplex PCR technique is useful for screening TCRBV usage and can be easily adapted for analysis of clonal composition in T-cell populations.

Original languageEnglish
Pages (from-to)122-134
Number of pages13
JournalTissue Antigens
Volume53
Issue number2
DOIs
Publication statusPublished - 01-03-1999
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Immunology
  • Cell Biology

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