To investigate T cell involvement in antigen-specific immune responses, it is important to detect accumulating T cells at a clonal level in vivo. However, thus far the clinical application of such analyses has been limited. Here we have established novel primers to anneal with T cell receptor (TCR) beta genes of multiple Vbeta families and applied them to reverse transcription-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis to evaluate peripheral T cell clonality of autoimmune disease patients. As a result, the new Vbeta primers could detect accumulating T cell clones in the periphery of healthy individuals and patients. It was revealed that patients with autoimmune diseases such as systemic lupus erythematosus (SLE) had a larger number of clonal accumulations of peripheral T cells compared with normal individuals. Thus, the RT-PCR-SSCP system using the new multifamily Vbeta primers is the first such laboratory examination to detect T cell clonal expansion, and will provide a simple and sensitive tool to aid in the diagnosis and also in the investigation of the pathogenesis of autoimmune diseases.