Hirose M, Abe-Hashimoto J, Tahara H, Ide T, Yoshimura T
1 Diagnostics Science Laboratory, Chugai Diagnostics Science Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171-8545, Japan.
Clin Chem. 1998 Dec;44(12):2446-52.
Telomerase is a ribonucleoprotein complex that uses RNA as a template for the addition of telomeric repeats. The development of the telomeric repeat amplification protocol (TRAP), a sensitive PCR-based assay, has facilitated the detection of telomerase activity in small tissue and tumor samples. Telomerase activity is expected to be a new diagnostic and prognostic marker of human cancer. In this study, we applied a non-PCR-based transcription-mediated amplification (TMA) and hybridization protection assay (HPA) to the measurement of telomerase activity by modification of both primers in TMA. We demonstrated that the modified TMA can detect and measure telomerase activity. TMA/HPA is as sensitive and reproducible as conventional TRAP, but is both faster and easier to perform. Furthermore, we found that TMA/HPA was influenced minimally by TRAP inhibitors that may come from clinical samples. TMA/HPA, which is easy, rapid, and applicable to a high-throughput format, should be clinically useful for the detection and monitoring of telomerase activity.
端粒酶是一种核糖核蛋白复合体，它以RNA为模板添加端粒重复序列。端粒重复序列扩增 protocol（TRAP）是一种基于PCR的灵敏检测方法，它的发展促进了在小组织和肿瘤样本中端粒酶活性的检测。端粒酶活性有望成为人类癌症的一种新的诊断和预后标志物。在本研究中，我们通过对TMA中的两种引物进行修饰，将基于非PCR的转录介导扩增（TMA）和杂交保护分析（HPA）应用于端粒酶活性的测量。我们证明了修饰后的TMA能够检测和测量端粒酶活性。TMA/HPA与传统的TRAP一样灵敏且可重复，但执行起来更快且更容易。此外，我们发现TMA/HPA受可能来自临床样本的TRAP抑制剂的影响最小。TMA/HPA简便、快速且适用于高通量形式，在临床上对于端粒酶活性的检测和监测应该是有用的。
