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

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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.

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