Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit
Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit
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DOI：
10.1007/bf00132889
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发表时间：
1996-01-01
期刊：
影响因子：
--
通讯作者：
Shay, Jerry W.
中科院分区：
文献类型：
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作者：
Holt, Shawn E.;Norton, James C.;Shay, Jerry W.
核糖核蛋白，端粒酶，被认为是负责维持不朽和癌细胞的端粒长度。建立了一种基于PCR的端粒酶活性检测方法(TRAP法：端粒重复序列扩增法)，可以在样本量有限的情况下快速有效地检测端粒酶活性。在使用TRAP试验检查的数千例原发人类肿瘤中，几乎90%的肿瘤显示出端粒酶活性。因此，为了癌症的早期发现和癌症治疗中化合物和药物的快速筛选，检测端粒酶活性的方法正在迅速出现。Oncor公司最近开发的TRAP-EZE试剂盒具有更高的灵敏度，同时减少了样品处理时间，从而改进了对大量样品中端粒酶活性的检测。在本研究中，我们使用细胞培养和临床材料解决了标准TRAP分析和TRAP-EZE试剂盒的重复性、可靠性和线性至关重要的一些技术方面和局限性。
The ribonucleoprotein, telomerase, is believed to be responsible for the maintenance of telomere length in immortal and cancer cells. A PCR-based assay for the detection of telomerase activity (TRAP assay: telomeric repeat amplification protocol) was developed, allowing fast and efficient detection of telomerase activity when sample amounts are limiting. Of the thousands of primary human tumors examined using the TRAP assay, almost 90% have been shown to exhibit telomerase activity. Thus, for the early detection of cancer and for the rapid screening of compounds and drugs in cancer therapeutics, methods for the detection of telomerase activity are rapidly emerging. The recently developed TRAP-eze kit from Oncor, Inc. gives increased sensitivity with decrease sample processing time, allowing improved detection of telomerase activity in a large number of samples. In the present study, we have addressed some of the technical aspects and limitations of critical importance for reproducibility, reliability, and linearity of the standard TRAP assay and the TRAP-eze kit using cell culture and clinical materials.