Rapid, nonradioactive detection of clonal T-cell receptor gene rearrangements in lymphoid neoplasms.
Southern blot hybridization analysis of clonal antigen receptor gene rearrangements has proved to be a valuable adjunct to conventional methods for diagnosing lymphoid neoplasia. However, Southern blot analysis suffers from a number of technical disadvantages, including the time necessary to obtain results, the use of radioactivity, and the susceptibility of the method to various artifacts. We have investigated an alternative approach for assessing the clonality of antigen receptor gene rearrangements in lymphoid tissue biopsy specimens. This approach involves the amplification of rearranged gamma T-cell receptor genes by the polymerase chain reaction and analysis of the polymerase chain reaction products by denaturing gradient gel electrophoresis. By use of this approach, clonal rearrangements from neoplastic lymphocytes constituting as little as 0.1-1% of the total cells in the tissue are detected as discrete bands in the denaturing gel after the gel is stained with ethidium bromide and viewed under ultraviolet light. In contrast, polyclonal rearrangements from reactive lymphocytes appear as a diffuse smear along the length of the gel. Our findings suggest that polymerase chain reaction combined with denaturing gradient gel electrophoresis may offer a rapid, nonradioactive, and sensitive alternative to Southern blot analysis for the diagnostic evaluation of lymphoid tissue biopsy specimens.
Bourguin
A
A
Department of Pathology, Brigham and Women's Hospital, Boston, MA.
Tung
R
R
Galili
N
N
Sklar
J
J
D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D002648:Child; D004247:DNA / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics / Q000276:immunology*; D008223:Lymphoma / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012016:Reference Values; D015183:Restriction Mapping; D013601:T-Lymphocytes / Q000276:immunology*
D004273:DNA, Neoplasm; D015345:Oligonucleotide Probes; D004247:DNA