Simple, reliable detection of T cell clones by PCR-LIS-SSCP analysis of TCRgamma rearrangement.
Clonal populations of T cells can be identified by polymerase chain reaction (PCR) amplification of the rearranged T cell receptor gamma (TCRG) chain gene. However, because of the limited combinatorial diversity of this locus it is necessary to separate the PCR product on the basis of sequence as well as size to distinguish clonal and polyclonal T cell populations. A simple method is described which achieves this by analysing the PCR product on a single-stranded conformation polymorphism (SSCP) gel. Sensitivity has been improved by denaturing the DNA using a low ionic strength (LIS) method rather than the more conventional alkali or formamide. Results from the PCR-LIS-SSCP method on a wide range of disorders and types of tissue samples show that clonality could be demonstrated in 40/44 cases.
Lynas
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Department of Haematology, Derriford Hospital, Plymouth, UK.
Howe
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D
D001483:Base Sequence; D002478:Cells, Cultured; D002999:Clone Cells; D017931:DNA Primers; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D006801:Humans; D007154:Immune System Diseases / Q000235:genetics / Q000276:immunology; D007938:Leukemia / Q000235:genetics / Q000276:immunology; D016399:Lymphoma, T-Cell / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
D017931:DNA Primers; D016692:Receptors, Antigen, T-Cell, gamma-delta