Evaluation of capillary electrophoresis in polymer solutions with laser-induced fluorescence detection for the automated detection of T-cell gene rearrangements in lymphoproliferative disorders

Robert P. Oda, Myra J. Wick, Lisa Marie Rueckert, John A. Lust, James P. Landers

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

The rapid increase in the number of DNA-based clinical diagnostic procedures, particularly polymerase chain reaction (PCR)-based assays, has generated interest in analytical techniques that are less time-consuming and labor-intensive than traditional procedures such as polyacrylamide gel electrophoresis (PAGE). Capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection, which allows for rapid and sensitive detection of DNA fragments in an automated format, is well-suited for DNA-based clinical assays. In this study, we demonstrate the potential of CE-LIF for the detection of PCR products from T-cell receptor gamma (TCRγ) gene rearrangements present in monoclonal populations of lymphocytes. The presence of monoclonal populations of T-cells is associated (but not always synonymous) with lymphocytic malignancies. Analysis of 31 patient samples, as well as sensitivity controls, demonstrated that CE-LIF detection of monoclonal lymphocytic populations is comparable to that of PAGE-SYBR Green I staining, and that CE-LIF detection can be accomplished in less than 20 min. These preliminary results illustrate the potential feasibility of a CE-based diagnostic assay for the detection of T-cell gene rearrangements.

Original languageEnglish (US)
Pages (from-to)1491-1498
Number of pages8
JournalElectrophoresis
Volume17
Issue number9
DOIs
StatePublished - Sep 1 1996

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Keywords

  • Capillary electrophoresis
  • Gene rearrangements
  • Laser-induced fluorescence
  • Polymer solutions
  • Polymerase chain reaction
  • T-cell receptor (TCRγ)

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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