Krupp G, Kühne K, Tamm S, Klapper W, Heidorn K, Rott A, Parwaresch R
Institut für Hämatopathologie, Christian-Albrechts-Universität Kiel, Kiel, Germany.
Nucleic Acids Res. 1997 Feb 15;25(4):919-21. doi: 10.1093/nar/25.4.919.
Human somatic cells have essentially no telomerase activity. Telomerase is linked to tumor genesis and is a valuable marker for malignant growth. Extreme paucity of the enzyme neccessitated development of a PCR-based assay, 'telomeric repeat amplification protocol' (TRAP). Unfortunately, this method is not without difficulties. Amplification products are not related to the size of the amplified telomerase products. Furthermore, false positive results can occur, and careful control of reaction conditions is crucial. We analyzed in detail the molecular basis of artifacts. Based on these data, reverse PCR primer was changed and both problems in the TRAP assay were eliminated.
人类体细胞基本上没有端粒酶活性。端粒酶与肿瘤发生有关，是恶性生长的一个重要标志物。由于该酶极其稀少，因此需要开发一种基于聚合酶链反应（PCR）的检测方法，即“端粒重复序列扩增 protocol”（TRAP）。不幸的是，这种方法并非没有困难。扩增产物与扩增的端粒酶产物大小无关。此外，可能会出现假阳性结果，因此仔细控制反应条件至关重要。我们详细分析了假象的分子基础。基于这些数据，改变了反向PCR引物，消除了TRAP检测中的两个问题。
