High-throughput ribozyme-based assays for detection of viral nucleic acids |
| |
Authors: | Kossen Karl Vaish Narendra K Jadhav Vasant R Pasko Christopher Wang Hong Jenison Robert McSwiggen James A Polisky Barry Seiwert Scott D |
| |
Institution: | Sirna Therapeutics Inc., 2950 Wilderness Place, Boulder, CO 80301 USA. |
| |
Abstract: | Many reports have suggested that target-activated ribozymes hold potential value as detection reagents. We show that a "half"-ribozyme ligase is activated similarly by three unstructured oligoribonucleotides representing the major sequence variants of a hepatitis C virus 5'-untranslated region (5'-UTR) target and by a structured RNA corresponding to the entire 5'-UTR. Half-ribozyme ligation product was detected both in an ELISA-like assay and in an optical immunoassay through the use of hapten-carrying substrate RNAs. Both assay formats afford a limit of detection of approximately 1 x 10(6) HCV molecules (1.6 attomol, 330 fM), a sensitivity which compares favorably to that provided by standard immunoassays. These data suggest that target-activated ribozyme systems are a viable approach for the sensitive detection of viral nucleic acids using high-throughput platforms. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|