Lynas C, Howe D
Department of Haematology, Derriford Hospital, Plymouth, UK.
Mol Cell Probes. 1998 Feb;12(1):41-8. doi: 10.1006/mcpr.1997.0146.
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.
T细胞的克隆群体可通过对重排的T细胞受体γ（TCRG）链基因进行聚合酶链反应（PCR）扩增来鉴定。然而，由于该基因座的组合多样性有限，有必要根据序列和大小对PCR产物进行分离，以区分克隆性和多克隆性T细胞群体。本文描述了一种简单的方法，通过在单链构象多态性（SSCP）凝胶上分析PCR产物来实现这一点。通过使用低离子强度（LIS）方法而非更传统的碱或甲酰胺对DNA进行变性，提高了灵敏度。PCR-LIS-SSCP方法对多种疾病和组织样本类型的检测结果表明，44例中有40例可证明存在克隆性。
