Journal
CHEMISTRY & BIOLOGY
Volume 11, Issue 6, Pages 807-815Publisher
CELL PRESS
DOI: 10.1016/j.chembiol.2004.03.029
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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.
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