Evaluation of capillary electrophoresis in polymer solutions with laser-induced fluorescence detection for the automated detection of T-cell gene rearrangements in 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.
Clinical Capillary Electrophoresis Facility, Mayo Foundation/Mayo Clinic, Rochester, MN 55905, USA.
R P
RP
Oda
M J
MJ
Wick
L M
LM
Rueckert
J A
JA
Lust
J P
JP
Landers
D001331:Automation; D052160:Benzothiazoles; D002999:Clone Cells; D003959:Diamines; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D004996:Ethidium; D005069:Evaluation Studies as Topic; D005240:Feasibility Studies; D005453:Fluorescence; D005456:Fluorescent Dyes; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D006801:Humans; D007834:Lasers; D008232:Lymphoproliferative Disorders / Q000235:genetics* / Q000473:pathology; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction; D011108:Polymers; D011804:Quinolines; D012680:Sensitivity and Specificity; D012996:Solutions
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011108:Polymers; D011804:Quinolines; D012996:Solutions; C098022:SYBR Green I; D004996:Ethidium