sybr-green-i and Lymphoproliferative-Disorders

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 (TCR gamma) 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.

Topics: Automation; Benzothiazoles; Clone Cells; Diamines; Electrophoresis, Capillary; Electrophoresis, Polyacrylamide Gel; Ethidium; Evaluation Studies as Topic; Feasibility Studies; Fluorescence; Fluorescent Dyes; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; Humans; Lasers; Lymphoproliferative Disorders; Organic Chemicals; Polymerase Chain Reaction; Polymers; Quinolines; Sensitivity and Specificity; Solutions